PROTEIN KINASES
SERINE/THREONINE KINASES
PROTEIN KINASE C
PKC ENZYMES
TABLE 1. PROTEIN KINASE C SUBTYPES
| Subtype |
MW (kDa) |
Activators |
Tissue Expression |
Refs. |
| Group A, Classical |
||||
| a |
76.8 |
PS, Ca2+, DG, FFA, LPC |
Ubiquitous |
1,2 |
| bI |
76.8 |
PS, Ca2+, DG, FFA, LPC |
Various |
1,2 |
| bII |
76.9 |
PS, Ca2+, DG, FFA, LPC |
Various |
1,2 |
| g |
78.4 |
PS, Ca2+, DG, FFA, LPC |
Brain |
1,2 |
| Group B, New |
||||
| d |
77.5 |
PS,DG |
Ubiquitous |
3,4,5 |
| e |
83.5 |
PS, DG, FFA |
Brain |
3,6,7,8 |
| h |
78.0 |
Cholesterol sulfate |
Lung, skin, heart |
9,10,11 |
| q |
81.6 |
? |
Skeletal muscle |
12 |
| Group C, Atypical |
||||
| z |
67.7 |
PS, FFA, PIP3, Cer |
Ubiquitous |
9,13,14 |
| l |
67.2 |
? |
Ovary, testis |
12 |
Abbreviations: DG, diacylglycerol; PS, phosphatidylserine; FFA, cis unsaturated fatty acid; LPC, lysophosphatidylcholine; Cer, ceramide. Adapted from reference 12.
SE-111
Protein kinase C, calcium- and lipid-dependent (from rat brain)
>95% by SDS-PAGE, MW=76-82 kDa (EC 2.7.1.37) Storage: -70°C
Supplied at 35 µg/ml. Specific activity: >1 µmol phosphate/min/mg enzyme using
histone H1 as substrate at pH 7.4 and 22°C. Consists primarily of a,
b and g isotypes15,16,17,18.
1.75 µg
SE-133
Protein kinase C, catalytic fragment (PKM) (from rat brain)
>95% by SDS-PAGE, MW=55 kDa, Storage: -70°C
Supplied at 10 µg/ml. Specific activity: 0.8 µmol phosphate/min/mg enzyme using histone
H1 as substrate at pH 7.4 and 30°C. This is the catalytically active fragment of rat
brain PKC (catalog #SE-111, above) prepared by digestion with trypsin19,20,21.
It does not require calcium or phosphatidylserine for its activity.
0.25 µg
5 x 0.25 µg
SE-143
Protein kinase C alpha (PKCa) (human, recombinant)
>95% by SDS-PAGE, MW=76.8 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.5 µmol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2,
100 µM CaCl2, 100 µg/ml phosphatidylserine (PS), 20 µg/ml
diacylglycerol (DAG), 0.2 mg/ml histone H1, 100 µM ATP and 0.03% Triton X-100 at pH 7.4
and 30°C. Activity of PKCa in the absence of PS and DAG is
decreased to <15%. May be used to phosphorylate target substrates of PKCa and to study enzyme regulation and kinetics.
10 µg
SE-144
Protein kinase C beta I (PKCbI) (human, recombinant)
>95% by SDS-PAGE, MW=76.8 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~0.9 µmol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2,
100 µM CaCl2, 100 µg/ml phosphatidylserine (PS), 20 µg/ml
diacylglycerol (DAG), 0.2 mg/ml histone H1, 100 µM ATP and 0.03% Triton X-100 at pH 7.4
and 30°C. Activity of PKCbI in the absence of PS and DAG is
decreased to <3%. May be used to phosphorylate target substrates of PKCbI and to study enzyme regulation and kinetics22,23,24.
10 µg
SE-146
Protein kinase C gamma (PKCg) (human, recombinant)
>95% by SDS-PAGE, MW=78.4 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 µmol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2, 100 µM
CaCl2, 100 µg/ml phosphatidylserine (PS), 20 µg/ml diacylglycerol (DAG), 0.2 mg/ml
histone H1, 100 µM ATP and 0.03% Triton X-100 at pH 7.4 and 30°C. Activity of PKCg in the absence of PS and DAG is decreased to <10%. May
be used to phosphorylate target substrates of PKCg and
to study enzyme regulation and kinetics.
10 µg
SE-147
Protein kinase C delta (PKCd) (human, recombinant)
>95% by SDS-PAGE, MW=77.5 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 µmol phosphate/min/mg protein
using PKC epsilon peptide (BIOMOL catalog #P-155) as substrate. Reaction conditions are 10
mM MgCl2, 0.1 mM EGTA, 200 µg/ml phosphatidylserine (PS), 20 µg/ml diacylglycerol (DAG),
0.1 mg/ml substrate, 100 µM ATP and 0.03% Triton X-100 at pH 7.4 and 30°C. May be used
to phosphorylate target substrates of PKCd and to study enzyme
regulation and kinetics24,25.
10 µg
SE-148
Protein kinase C epsilon (PKCe) (human, recombinant)
>95% by SDS-PAGE, MW=83.5 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.1 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 µmol phosphate/min/mg protein
using PKC epsilon peptide (BIOMOL catalog #P-155) as substrate. Reaction conditions are 10
mM MgCl2, 0.1 mM EGTA, 200 µg/ml phosphatidylserine (PS), 20 µg/ml
diacylglycerol (DAG), 0.1 mg/ml substrate, 100 µM ATP and 0.03% Triton X-100 at pH 7.4
and 30°C. May be used to phosphorylate target substrates of PKCe
and to study enzyme regulation and kinetics24,25.
10 µg
SE-149
Protein kinase C zeta (PKCz) (human, recombinant)
>95% by SDS-PAGE, MW=67.7 kDa, (EC 2.7.1.37) Storage: -70°C
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.2 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~0.4 µmol phosphate/min/mg protein
using PKC epsilon peptide (BIOMOL catalog #P-155) as substrate. Reaction conditions are 5
mM MgCl2, 0.5 mM EGTA, 1 mM DTT, 100 µg/ml phosphatidylserine (PS), 0.1 mg/ml
substrate, and 100 µM ATP at pH 7.5 and 30°C. May be used to phosphorylate target
substrates of PKCz and to study enzyme regulation and
kinetics23,24
10 µg
PKC SUBSTRATES
P-117
MARCKS psd peptide, PKC substrate
Lys-Lys-Lys-Lys-Lys-Arg-Phe-Ser-Phe-Lys-Lys-Ser-Phe-Lys-Leu-Ser-Gly-Phe-Ser-Phe-Lys-Lys-Asn-Lys-Lys
>97%, MW=3078.8 [125267-21-2] Storage: -20°C
Sequence is from myristoylated Ala-rich C-kinase substrate (MARCKS) protein phosphorylated
site domain (psd), residues 151-175. Protein kinase C (including PKCe)
phosphorylates the peptide at Ser-8, 12 and 19. Poor substrate for PKA, PKG, CaMKI, II,
III. Km=20 nM for purified rat brain PKC. Can be used in phosphocellulose,
SDS-PAGE or anion exchange assays26,27,28.
1 mg
5 x 1 mg
P-155
Protein kinase C epsilon substrate
Glu-Arg-Met-Arg-Pro-Arg-Lys-Arg-Gln-Gly-Ser-Val-Arg-Arg-Arg-Val
>97%, MW=2066.2, Storage: -20°C
PKC epsilson pseudosubstrate sequence (149-164) but Ala to Ser-159 substitution makes it a
substrate for phosphorylation29. Phospholipid/phorbol
ester-dependent substrate for PKC e and z 6,14.
0.5 mg
5 x 0.5 mg
P-110
MBP (4-14), N-acetylated, protein kinase C substrate
N-Acetyl-Gln-Lys-Arg-Pro-Ser-Gln-Arg-Ser-Lys-Tyr-Leu
>97%, MW=1432.8 [126768-94-3] Storage: -20°C
Sequence is from myelin basic protein (4-14)30. Specific
substrate for a-,b-, g-isoforms of PKC (Km=7 µM) and poor substrate for PKA,
CaMKII, CKI, CKII, and phosphorylase kinase. This N-acetylated peptide is resistant to
phosphatases in brain hippocampal extracts and therefore is a good substrate for use in
crude extracts31.
1 mg
5 x 1 mg
P-109
EGF-R fragment (651-658), protein kinase substrate
Arg-Lys-Arg-Thr-Leu-Arg-Arg-Leu
>97%, MW=1098.5, Storage: -20°C
Epidermal growth-factor receptor (EGF-R) conserved sequence which is identical to v-erb-B
(95-102). Sequence includes EGF-R Thr-654 PKC phosphorylation site32.
1 mg
5 x 1 mg
P-111
Protein kinase C [Ser-25] (19-31), substrate
Arg-Phe-Ala-Arg-Lys-Gly-Ser-Leu-Arg-Gln-Lys-Asn-Val
>97%, MW=1560, Storage: -20°C
PKC pseudosubstrate sequence but Ala to Ser-25 substitution makes it a good substrate (Km=0.2
µM) rather than an inhibitor33,34.
1 mg
5 x 1 mg
PKC ACTIVATORS
PHORBOL ESTERS
Phorbol esters such as phorbol 12-myristate 13-acetate (PMA), phorbol 12,13-dibutyrate
(PDBu), and phorbol 12,13-didecanoate (PDD) bind to and activate protein kinase C. At
nanomolar concentrations, they dramatically increase the affinity of the enzyme for Ca2+
resulting in its full activation35-39. Phorbol esters with 4a stereochemistry are inactive and may be used as negative controls40.
WARNING! Potential health hazard: Phorbol esters are vesicants. Irritation and blistering
may result from skin contact. Use of these compounds should be restricted to technically
qualified personnel.
PE-187
12-Deoxyphorbol 13-acetate (Prostratin)
99%, MW=390.5 [60857-08-1] Storage: -20°C
A non-tumor promoting phorbol ester which acts as a PKC partial agonist. Protects
T-lymphoblastoid cells from the killing effects of HIV-141.
1 mg
5 mg
PE-182
12-Deoxyphorbol 13-phenylacetate 20-acetate (dPPA)
99%, MW=508.6 [54662-30-5] Storage: -20°C
dPPA is a selective activator of PKCb1. It is at least
1000-fold more potent for the b1 isozyme than the a, g, d or e isozymes but activates the isozyme to only 60% of maximal activity42,43. Mimicks the effect of TNFa on
endothelial cells but does not translocate PKC44.
1 mg
5 mg
PE-135
Phorbol 12,13-dibutyrate
99%, MW=504.6 [37558-16-0] Storage: -20°C
PKC activator35 (10-100 ng/ml).
1 mg
5 mg
PE-140
4a-Phorbol 12,13-dibutyrate
99%, MW=504.6 [93781-54-5] Storage: -20°C
Negative control for phorbol 12,13-dibutyrate.
1 mg
5 mg
PE-145
Phorbol 12,13-didecanoate
99%, MW=672.9 [24928-17-4] Storage: -20°C
PKC activator35 (10-100 ng/ml).
1 mg
5 mg
PE-147
4a-Phorbol 12,13-didecanoate
99%, MW=672.9 [27536-56-7] Storage: -20°C
Negative control for phorbol 12,13-didecanoate35.
1 mg
5 mg
PE-160
Phorbol 12-myristate 13-acetate (PMA)
(12-tetradecanoyl phorbol 13-acetate, TPA)
99%, MW=616.8 [16561-29-8] Storage: -20°C
PKC activator35,42 (10-100 ng/ml).
1 mg
5 mg
PE-162
4a-Phorbol 12-myristate 13-acetate
99%, MW=616.8 [63597-44-4] Storage: -20°C
Negative control for phorbol 12-myristate 13-acetate (PMA).
1 mg
5 mg
DIACYLGLYCEROLS AND RELATED PRODUCTS
A variety of natural and unnatural diacylglycerols (DAG) bind to and activate protein
kinase C38,45,46. Activation by diacylglycerols is transient
due to their rapid conversion by diacylglycerol kinase47 and
lipase48. Diacylglycerols with various fatty acids in the
1,2-sn configuration are active, with those having an unsaturated fatty acid being most
active45,49,50. The other isomers, 2,3-sn-diacylglycerol and
l,3-diacylglycerol, neither activate nor inhibit the enzyme51.
Activation of protein kinase C by diacylglycerols is synergistic with cis-unsaturated
fatty acids52. Diacylglycerols play important roles in
growth factor signal transduction53.
SE-100
Diacylglycerol kinase (from E. coli)
sn-1,2-diacylglycerol kinase (DAGK)
MW=13.7 kDa, [93076-89-2] (EC 2.7.1.107) Storage: -20°C
Supplied as a turbid membrane suspension with protein concentration of 1 mg/ml. Specific
for sn-1,2 isomer of DAG but will phosphorylate ceramide and monoglycerol at a lower rate.
Specific activity is >5 U/mg protein for DAG. One unit is the amount of enzyme that
will phosphorylate 1.0 mmol of diacylglycerol per minute at
25°C54. Specific activity for ceramide phosphorylation is
~0.5 mmol ceramide/min/mg protein at 25°C. Can be used to
quantify diacylglycerol and ceramide in crude lipid extracts55,56.
1 mg
5 x 1 mg
DG-100
(?1,2-Didecanoylglycerol (10:0)
98%, MW=400.6 [82950-64-9] Storage: -20°C
A membrane permeable but non-physiologic DAG57. Stimulates
protein kinase C in intact platelets50.
20 mg
100 mg
DG-112
1,2-Dioctanoyl-sn-glycerol (8:0)
98%, MW=344.5, [75685-80-2] Storage: -20°C
A membrane permeable but non-physiologic DAG50. Induces a
discrete but transient translocation of protein kinase C58.
20 mg
100 mg
DG-115
(?1,2-Dioleoylglycerol (18:1)
98%, MW=621.0 [3738-74-7] Storage: -20°C
PKC activator.
20 mg
100 mg
DG-125
1-Oleoyl-2-acetyl-sn-glycerol (OAG)
98%, MW=398.6 [86390-77-4] Storage: -20°C
A potent, membrane permeable, non-physiologic DAG which activates protein kinase C45,46.
25 mg
100 mg
DG-130
1-Stearoyl-2-arachidonoyl-sn-glycerol
98%, MW=645.0, [65914-84-3] Storage: -20°C
Putative endogenous activator of protein kinase C produced via receptor mediated inositol
phospholipid hydrolysis50.
10 mg
50 mg
DG-135
1-Stearoyl-2-linoleoyl-sn-glycerol
98%, MW=621.0, Storage: -20°C
Putative endogenous activator of protein kinase C produced via receptor mediated inositol
phospholipid hydrolysis50.
20 mg
100 mg
DG-140
1-O-Hexadecyl-2-O-arachidonoyl-sn-glycerol
98%, MW=603.0, Storage: -20°C
Hexadecylether analog of DG-130.
5 mg
25 mg
EI-136
R 59022
99%, MW=459.9 [93076-89-2] Storage: RT
R 59022 amplifies protein kinase C activity via inhibition of diacylglycerol kinase. It
inhibits human platelet DAG kinase in intact cells (IC50=3.6 µM) and in
isolated membranes (IC50=6.7 µM)59,60. In
porcine thymus cytosol it inhibited the 80 kDa DAG kinase (IC50=10 µM) with no
effect on the 150 kDa DAG kinase61.
10 mg
50 mg
EI-202
R 59949
99%, MW=489.6 [120166-69-0] Storage: 0°C
R 59949 is a more potent analog of R 59022. It inhibits human platelet DAG kinase in
intact cells (IC50=0.12 µM) and in isolated membranes (IC50=0.3
µM)62.
10 mg
50 mg
ST-300
RHC-80267
Diacylglycerol lipase inhibitor.
OTHER PKC ACTIVATORS
ST-103
Bryostatin 1
99%, MW=887.0 [83314-01-6] Storage: -20°C
A structurally unique marine natural product that binds to and activates PKC at picomolar
concentrations but is not a carcinogen or a complete tumor promoter63.
In many systems it induces only a subset of the responses to PMA and blocks those which it
does not induce64. It is markedly more potent than PMA for
translocating PKCd and e64.
10 µg
5 x 10 µg
PE-190
Farnesyl thiotriazole
98%, MW=305.5 [156604-45-4] Storage: -20°C
A structurally unique activator of PKC in whole cells or in purified enzyme preparations
(KD=2.5 µM) which is equipotent with dioleylglycerol65.
Since it does not possess hydrolyzable esters of fatty acids, it represents a new stable,
persistent PKC activator.
10 mg
50 mg
PE-186
Ingenol 3,20-dibenzoate
97%, MW=556.7 [59086-90-7] Storage: -20°C
Ingenol is a diterpenoid related to phorbol. Ingenol 3,20-dibenzoate competes with [3H]phorbol
dibutyrate for binding to PKCa (Ki for binding=240
nM)66.
1 mg
5 mg
PE-174
(-)-7-Octylindolactam V
99%, MW=413.6 [109346-66-9] Storage: -20°C
Octylindolactam V is a non-phorbol protein kinase C activator related to teleocidin. The
advantages of octylindolactam V include greater metabolic stability and high potency67,68 (inhibition of PMA binding: EC50=29 nM).
1 mg
5 mg
PE-176
(+)-7-Octylindolactam V
99%, MW=413.6 [123597-54-6] Storage: -20°C
A biologically inactive isomer which may be used as a negative control.
1 mg
5 mg
EI-143
SC-10
N-Heptyl-5-chloronaphthalene-1-sulfonamide
99%, MW=339.9 [102649-79-6] Storage: RT
Activates protein kinase C. Stimulates protein kinase C mediated myosin light chain
phosphorylation without activating MLC kinase69.
10 mg
50 mg
PE-184
Thymeleatoxin
99%, MW=628.7, Storage: -20°C
Thymeleatoxin is a diterpene isolated from Thymelea hirsuta L. It activates PKC
isozymes a, bI and g selectively (ED50 100 nM) over the d
and e isozymes42,43.
1 mg
5 mg
PKC INHIBITORS
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES
ISOTYPE-SELECTIVE PKC INHIBITORS
EI-269
G?6976
95%, MW=378.4 Storage: -20?/font>C
G?6976 inhibits PKCa (IC50=2.3 nM) and PKCb1 (IC50=6.2 nM) but has no effect on d,
e or z isotypes70.
500 mg
5 x 500 mg
EI-270
Rottlerin
98%, MW=516.6 [82-08-6] Storage: -20?/font>C
Inhibits PKCd (IC50=3-6 mM)
5-10 fold more potently than a and b
and 13-33 fold more potently than e, z
and h71.
10 mg
50 mg
EI-273
HBDDE
2,2',3,3'4,4'-Hexahydroxy-1,1'-biphenyl-6,6'-dimethanol dimethyl ether
97%, MW=338.4 Storage: -20?/font>C
Inhibits PKCa (IC50=43 mM)
and PKCg (IC50=50 mM)
without inhibiting the d, bI and bII isotypes72.
1 mg
5 mg
PAN-SPECIFIC PKC INHIBITORS
EI-198
Calphostin C (UCN-1028C)
99%, MW=790.6 [121263-19-2] Storage: 0°C
Calphostin C is a novel, potent and highly selective inhibitor of protein kinase C (IC50=0.05
µM)73,74. Inhibition is via interaction with the regulatory
domain75 and is light dependent76.
See Table 4 for selectivity data.
100 µg
1 mg
EI-225
Chelerythrine chloride
98%, MW=383.8 [3895-92-9] Storage: 0°C
A potent and selective inhibitor of protein kinase C77 (IC50=0.66
µM). See Table 4 for selectivity data.
5 mg
25 mg
EI-246
GF 109203X
98%, MW=412.5 [133052-90-1] Storage -20°C
GF 109203X is a potent and selective protein kinase C inhibitor which has been used
effectively in platelets, Swiss 3T3 fibroblasts78
and macrophages79. Inhibitory profile: PKC, IC50=0.02
µM (inhibits a, bI, bII and g subtypes with similar potency);
PKA, IC50=2.0 µM; phosphorylase kinase, IC50=0.7 µM; insulin, EGF
and PDGF receptor tyrosine kinases are not inhibited at concentrations up to 50 µM78.
1 mg
5 x 1 mg
EI-149
(?1-O-Hexadecyl-2-O-methylglycerol
99%, MW=330.6, Storage: 0°C
Inhibits protein kinase C activation by diacylglycerols80,81.
Inhibits the fMLP or phorbol ester-induced respiratory burst in human neutrophils82.
50 mg
250 mg
EI-150
1-O-Hexadecyl-2-O-acetyl-sn-glycerol
Inhibits protein kinase C activation by diacylglycerols.
EI-226
Hypericin
99%, MW=504.4 [548-04-9] Storage: 0°C
A potent and selective inhibitor of protein kinase C136 (IC50=3.4
µM) which also possesses antiviral activity83. See Table 4
for selectivity data.
1 mg
5 mg
EI-154
Phloretin
99%, MW=274.3 [60-82-2] Storage: 0°C
Inhibits protein kinase C. Completely inhibits transformation of BALB/3T3 cells at 30
µg/ml84.
200 mg
1 g
EI-155
Sphingosine
Inhibits protein kinase C activity and phorbol dibutyrate binding in human platelets.
See Sphingolipids
P-205
Myristoylated protein kinase C (18-28), cell-permeable PKC inhibitor
N-Myr-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln
>97%, MW=1256.0, Storage: -20°C
Pseudosubstrate sequence from PKCa and b.
Highly specific inhibitor which is N-terminal myristoylated to allow membrane
permeability. IC50=8 µM for TPA activation of MARCKS phosphorylation in
fibroblast primary cultures, and 98% inhibition at 100 µM85.
Partially reverses multidrug resistance in human breast cancer cells86.
0.5 mg
5 x 0.5 mg
P-207
Myristoylated EGF-R fragment (651-658), PKC inhibitor
N-Myr-Arg-Lys-Arg-Thr-Leu-Arg-Arg-Leu
>97%, MW=1308.1, Storage: -20°C
Epidermal growth-factor receptor (EGF-R) conserved sequence which is identical to v-erb-B
(95-102). This inhibitor is N-terminal myristoylated to allow membrane permeability and
inhibits protein kinase C in intact cells (IC50=5 µM)87,88.
0.5 mg
5 x 0.5 mg
P-206
Protein kinase C (19-31), pseudosubstrate inhibitor
Arg-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln-Lys-Asn-Val
>97%, MW=1544, Storage: -20°C
Shorter and more potent inhibitor of PKC (IC50=100 nM) than longer sequence
(19-36)33.
1 mg
5 x 1 mg
P-150
Protein kinase C pseudosubstrate (19-36), inhibitor
Arg-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln-Lys-Asn-Val-His-Glu-Val-Lys-Asn
>97%, MW=2151.8, Storage: -20°C
Corresponds to PKC pseudosubstrate regulatory sequence89.
Potent and selective PKC inhibitor which binds to the active site of the kinase. IC50
for purified brain PKC=0.18 µM, PKA=423 µM, MLCK=24 µM33,34.
0.5 mg
5 x 0.5 mg
PKC ANTIBODIES
SA-143
Anti-Protein kinase C
Mouse monoclonal antibody. Clone MC5. Supplied as affinity purified IgG2a, 0.1 mg/ml.
Recognizes Ca2+/phospholipid dependent protein kinase C (a,
b and g isotypes) in human, rat and
mouse. Epitope is at the hinge region of the enzyme, the site of phorbol ester induced
cleavage90. Applications: WB 1/100, IF 1/10 and other
techniques.
100 µg
SA-144
Anti-Protein kinase C a
Rabbit polyclonal antibody. Immunogen: Rabbit PKC a
(662-672) peptide. Supplied as serum. Cross-reacts with human, rat, mouse and other
mammalian species. Applications: WB 1/100.
1 ml
SP-144
Anti-Protein kinase C a (SA-144) control peptide
Control peptide CQFVHPILQSSV is useful to block specific staining by WB with SA-144,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-145
Anti-Protein kinase C b1
Rabbit polyclonal antibody. Immunogen: Human PKC bI (661-671)
peptide. Supplied as serum. Cross-reacts with rat, mouse and other mammalian species.
Applications: WB 1/100.
1 ml
SP-145
Anti-Protein kinase C b1 (SA-145) control peptide
Control peptide CSYTNPEFVINV is useful to block specific staining by WB with SA-145,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-146
Anti-Protein kinase C bII
Rabbit polyclonal antibody. Immunogen: Human PKC b II
(661-673) peptide. Supplied as serum. Cross-reacts with rat, mouse and other mammalian
species. Applications: WB 1/100.
1 ml
SP-146
Anti-Protein kinase C bII (SA-146) control peptide
Control peptide CFVNSEFLKPEVKS is useful to block specific staining by WB with SA-146,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-147
Anti-Protein kinase C g
Rabbit polyclonal antibody. Immunogen: Human PKC g
(681-697) peptide. Supplied as serum. Cross-reacts with rat, mouse and other mammalian
species. Applications: WB 1/100.
1 ml
SP-147
Anti-Protein kinase C g (SA-147) control peptide
Control peptide CVHPDARSPISPTPVPVM is useful to block specific staining by WB with SA-147,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-148
Anti-Protein kinase C d
Rabbit polyclonal antibody. Immunogen: Human PKC d
(658-676) peptide. Supplied as serum. Cross-reacts with rat, mouse and other mammalian
species. Applications: WB 1/100.
1 ml
SP-148
Anti-Protein kinase C d (SA-148) control peptide
Control Peptide CSAFAGFSFVNP is useful to block specific staining by WB with SA-148,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-149
Anti-Protein kinase C e
Rabbit polyclonal antibody. Immunogen: Human PKC e (721-737)
peptide. Supplied as serum. Cross-reacts with rat, mouse and other mammalian species.
Applications: WB 1/100.
1 ml
SP-149
Anti-Protein kinase C e (SA-149) contol peptide
Control peptide CNQEEFKFSYFGED is useful to block specific staining by WB with SA-149,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-150
Anti-Protein kinase C
Rabbit polyclonal antibody. Immunogen: Human PKC (721-737) peptide. Supplied as serum.
Cross-reacts with rat, mouse and other mammalian species. Applications: WB 1/100.
1 ml
SP-150
Anti-Protein kinase C (SA-150) contol peptide
Control peptide CFEGFEYINPLLLSTEESV is useful to block specific staining by WB with
SA-150, above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-151
Anti-Protein kinase C
Rabbit polyclonal antibody. Immunogen: Human PKC (666-683) peptide. Supplied as serum.
Cross-reacts with rat, mouse and other mammalian species. Applications: WB 1/100.
1 ml
SP-151
Anti-Protein kinase C (SA-151) control peptide
Control peptide CNQDEFRNFSYVSPELQP is useful to block specific staining by WB with SA-151,
above. Supplied at 0.2 mg/ml in ammonium bicarbonate buffer.
100 µg
SA-183
Anti-Protein kinase C set
Contains 0.5 ml each of SA-144, SA-145, SA-146, SA-147, SA-148, SA-149, SA-150, SA-151.
1 set
SP-183
PKC peptide set
Contains 100 µg each of SP-144, SP-145, SP-146, SP-147, SP-148, SP-149, SP-150, and
SP-151
1 set
PROTEIN KINASE A (PKA)
PKA ENZYMES
SE-110
Protein kinase A, a catalytic subunit (from porcine heart)
3',5'-cyclic AMP dependent protein kinase
MW=38 kDa, Storage: -20°C
Partially purified preparation, supplied lyophilized. Specific activity: 30-65 U/µg
protein. One unit will add 1 pmol phosphate to dephosphorylated casein per minute at pH
6.5 and 30°C. cAMP is not necessary for activity. Useful to phosphorylate peptides for
use as substrates in Ser/Thr phosphatase assays 92,94.
1000 U
SE-122
Protein kinase A, a catalytic subunit (murine, recombinant)
3',5'-cyclic AMP dependent protein kinase
MW=38 kDa, Storage: -20°C
Expressed in E. coli91. Contains no detectable
protease or phosphatase activity. Supplied at ~1 mg/ml in 50% glycerol buffer. Specific
activity: >1000 U/mg protein. One unit will add 1 nmol phosphate to Kemptide peptide
substrate (catalog #P-107, below) per minute at pH 7.5 and 30°C. Regulatory subunit is
not present, therefore cAMP is not necessary for activity. Useful to phosphorylate target
regulatory enzymes and other substrates of PKA.
50 U
5 x 50 U
TABLE 2. PROTEIN KINASE A ACTIVATORS: CYCLIC AMP ANALOGS*
| Catalog # |
Analog |
Comments |
Page # |
| CN-115 |
8-Bromo-cAMP |
Activates PKA, more resistant to PDE's than cAMP. |
000 |
| CN-136 |
Sp-cAMPS |
Activates PKA, completely resistant to PDE's, half-maximal activation at
1.8 µM. |
000 |
| CN-130 |
8-CPT cAMP |
Activates PKA and PKG. |
000 |
| CN-125 |
Dibutyryl-cAMP |
Preferentially activates PKA. |
000 |
| CN-120 |
Sp-5,6-DCl-cBiMPS |
Specific activator of PKA, resistant to PDE's. |
000 |
* All of these reagents are cell-permeable.
Abbreviations: PDE's = phosphodiesterases.
SEE CYCLIC AMP ANALOGS, ADENYLATE CYCLASE ACTIVATORS AND INHIBITORS AND PHOSPODIESTERASE
INHIBITORS FOR OTHER PKA RELATED REAGENTS.
PKA SUBSTRATES
P-118
[Ala-97]-RII (81-99), PKA substrate
Asp-Leu-Asp-Val-Pro-Ile-Pro-Gly-Arg-Phe-Asp-Arg-Arg-Val-Ser-Val-Ala-Ala-Glu
>97%, MW=2112.4, Storage: -20°C
Sequence is from cAMP-dependent protein kinase regulatory subunit type II (PKA). The
phosphorylated peptide can be used as a substrate for calcineurin92,93,94.
0.5 mg
5 x 0.5 mg
P-106
H1-7 (histone H1 phosphorylation site), PKA substrate
Arg-Arg-Lys-Ala-Ser-Gly-Pro
>97%, MW=770.9 [65189-70-0], Storage: -20°C
Sequence is derived from calf thymus histone H1 containing Ser-38. Peptide is
phosphorylated by protein kinase A95.
1 mg
5 x 1 mg
P-107
Kemptide, protein kinase substrate
Leu-Arg-Arg-Ala-Ser-Leu-Gly
>97%, MW=771.9 [65189-71-1] Storage: -20°C
Substrate for Ser/Thr protein kinases. Km=4.7 µM for PKA96,97,98.
1 mg
5 x 1 mg
P-108
Malantide, protein kinase substrate
Arg-Thr-Lys-Arg-Ser-Gly-Ser-Val-Tyr-Glu-Pro-Leu-Lys-Ile
>97%, MW=1634.1 [86555-35-3] Storage: -20°C
Sequence is from site of phosphorylation by PKA on the b-subunit
of phosphorylase kinase. Highly specific substrate for cAMP-dependent protein kinase (PKA)99. PKI inhibited >90% substrate phosphorylation in various
tissue extracts100. Km=15 µM for PKA; 223 µM
for PKG.
1 mg
5 x 1 mg
PKA INHIBITORS
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES)
EI-196
H-89
N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide?HCl
99%, MW=519.2 [127243-85-0] Storage: 0°C
H-89 is a potent, selective101 and widely used102,103 inhibitor of PKA (Ki=48 nM). See Table 4
for selectivity data.
5 mg
25 mg
EI-199
KT5720
98%, MW=537.6 [108068-98-0] Storage: 0°C
KT5720 is a hexylester derivative of K-252a and is a selective inhibitor of PKA (Ki=0.056
µM)193,104. See Table 4 for
selectivity data.
100 µg
5 x 100 µg
EI-158
H-8
N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide?HCl
99%, MW=338.3 [84478-11-5] Storage: 0°C
H-8 inhibits PKA and PKG selectively over PKC and MLCK190,105. See Table 4 for selectivity data.
10 mg
50 mg
EI-184
HA-1004
N-(2-Guanidinoethyl)-5-isoquinolinesulfonamide?HCl
98%, MW=329.8 [91742-10-8] Storage: 0°C
HA-1004 inhibits PKA and PKG selectively over PKC and MLCK190.
See Table 4 for selectivity data. May be used in conjunction with H7 to probe for
PKC-specific effects106.
10 mg
50 mg
EI-233
HA-1077
1-(5-Isoquinolinesulfonyl)-homopiperazine·HCl
98%, MW=327.8 [103745-39-7] Storage: 0°C
HA-1077 inhibits PKA and PKG selectively over MLCK107,108.
See Table 4 for selectivity data. Potent vasodilator109.
10 mg
50 mg
SE-123
PKI, Heat-stable inhibitor of cAMP-dependent protein kinase (Rabbit muscle, recombinant)
MW=8 kDa, Storage: -20°C
Native protein (77 amino acid) with an additional N-terminal Met and Gly, expressed in E.
coli110. Contains no detectable protease or phosphatase
activity. Supplied at 0.8 mg/ml in 50% glycerol buffer. Specific activity: 2250 U/mg
protein. One unit will inhibit 1 unit of PKA activity as defined by product SE-122. Ki
for PKA catalytic subunit = 0.1 nM. Selective for PKA111.
50 U
5 x 50 U
P-203
PKI (5-24), cAMP-dependent protein kinase inhibitor
Thr-Thr-Tyr-Ala-Asp-Phe-Ile-Ala-Ser-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile-His-Asp
>97%, MW=2222.7 [99534-03-9] Storage: -20°C
Sequence is from the heat-stable skeletal muscle inhibitor protein of PKA. Peptide binds
to the catalytic subunit of PKA and displaces the regulatory subunit. Mimics protein
substrate by binding to the catalytic site via the Arg-cluster basic residues. Ki
=2.3 nM for PKA112,113.
0.5 mg
5 x 0.5 mg
P-204
PKI(6-22) amide, cAMP-dependent protein kinase inhibitor
Thr-Tyr-Ala-Asp-Phe-Ile-Ala-Ser-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile-NH2
>97%, MW=1868.1 [121932-06-7] Storage: -20°C
Competitive inhibitor of PKA (Ki =1.7 nM)114,115.
1 mg
5 x 1 mg
P-210
Myristoylated PKI (14-22) amide, cell-permeable PKA inhibitor
>97%, MW=1208.9 Storage: -20°C
Heat-stable protein kinase inhibitor (PKI) peptide sequence (14-22)115.
N-terminal is myristoylated to increase cell membrane permeability85,88.
The non-myristoylated version of this peptide is a highly specific inhibitor of
cAMP-dependent protein kinase (PKA) with Ki=36 nM115.
This product is a new useful tool for studying PKA in cellular systems.
0.5 mg
5 x 0.5 mg
CN-135
Rp-cAMPS
Rp-Adenosine-3',5'-cyclic monophosphorothioate (Rp-cAMPS)
Rp-cAMPS is a potent and specific competitive inhibitor of the activation of cAMP
dependent protein kinases by cAMP116 (IC50=4.9
µM)117.
PKA ANTIBODIES
SA-195
Anti-Protein kinase A, RII subunit
Mouse monoclonal antibody. Clone R-30. Immunogen: Purified bovine brain regulatory subunit
RII of cAMP-dependent protein kinase118. Supplied as
affinity purified IgG2a, 1 mg/ml. Epitope is N-terminal non-conserved region of RII
(1-91). Cross-reacts with porcine and to a lesser extent human, rat, and mouse. Antibody
enhances binding of RII for cAMP, and does not interfere with the association of RII and C
subunits. Applications: WB, IF, ELISA.
250 µg
SW-107
Bovine Heart Extract, Immunoblotting standard
Heart extract purified from cow. Supplied as tissue extract (approximately 0.5 mg/ml) in
SDS-PAGE sample buffer, ready-to-use at 2-5 µl (~1-2.5 µg) per lane. Useful as a Western
blotting control for anti-protein kinase A, RII subunit (SA-195, above), or for use with
other antibodies.
200 µl
SA-270
Anti-Protein kinase A, RIIb subunit (PKA, RIIb)
Rabbit polyclonal antibody. Immunogen: Mouse protein kinase A, regulatory subunit RIIb, N-terminal peptide conjugated to KLH. Supplied as serum.
Recognizes human, rat and mouse 52 kDa PKA RIIb. It
does not crossreact with PKA RIa, RIb, or RIIa. Applications: WB
1/1000 (alkaline phosphatase), IF and IP119,120.
100 µl
SA-271
Anti-Protein kinase A, RIIb subunit (PKA, RIIb), purified
Rabbit polyclonal antibody. Identical to SA-270 above, but supplied as protein A purified
IgG.
100 µg
SP-270
Anti-PKA RIIb (SA-270, -271) control peptide
>90%, MW=1980.0 Storage: -20°C
Immunization and blocking peptide for SA-270 and -271 above. Useful to block specific
staining by WB with anti-PKA RIIb antibody (SA-270 and -271,
above). Supplied at 1 mg/ml in PBS.
100 µg
SW-103
Rat Brain Extract, Immunoblotting Standard
Whole brain protein extract from rat. Supplied as tissue extract (approximately 5 mg/ml)
in SDS-PAGE sample buffer, ready-to-use at 5 µl/lane. Useful as a Western blotting
control for PKA RIIb antibody (SA-270 and -271, above), or for
use with other antibodies.
200 µl
SW-104
Mouse Brain Extract, Immunoblotting Standard
Whole brain protein extract from mouse. Similar to SW-103 above, but from mouse.
200 µl
PROTEIN KINASE G (PKG)
PKG ENZYME
SE-118
Protein kinase G (from bovine lung)
cGMP-dependent protein kinase
90%, Storage: -70°C
Specific activity: 1-2.5 kU/µg protein, 10-20 pmol cGMP bound/µg. One unit will transfer
1 pmol phosphate to RKRSRAE peptide substrate per minute122,121.
6 kU
5 x 6 kU
TABLE 3. PROTEIN KINASE G ACTIVATORS: CYCLIC GMP ANALOGS*
| Catalog # |
Analog |
Comments |
Page # |
| CN-206 |
Rp-8-pCPT-cGMPS |
PKG antagonist (Ki=0.5 µM), and is resistant to PDE's. |
000 |
| CN-216 |
Rp-8-Br-cGMPS |
PKG antagonist, and is resistant to PDE's. |
000 |
| CN-205 |
8-Bromo-cGMP |
PKG activator, more resistant to PDE's than cGMP. |
000 |
| CN-207 |
Sp-8-pCPT-cGMPS |
Activates PKA and PKG; cGMP mimetic. |
000 |
| CN-215 |
Dibutyryl-cGMP |
Preferentially activates PKG. |
000 |
| CN-120 |
Sp-5,6-DCl-cBiMPS |
Specific activator of PKA, resistant to PDE's. |
000 |
| CN-217 |
Sp-8-Br-cGMPS |
Activates PKA and PKG; cGMP mimetic. |
000 |
* All of these reagents are cell-permeable.
Abbreviations: PDE's = phosphodiesterases.
SEE CYCLIC GMP ANALOGS, GUANYLATE CYCLASE ACTIVATORS AND INHIBITORS AND PHOSPODIESTERASE
INHIBITORS FOR OTHER PKG RELATED REAGENTS.
PKG SUBSTRATE
P-112
BPDEtide, cGMP-dependent protein kinase (PKG) substrate
Arg-Lys-Ile-Ser-Ala-Ser-Glu-Phe-Asp-Arg-Pro-Leu-Arg
>97%, MW=1575, Storage: -20°C
Sequence derived from bovine lung cGMP specific phosphodiesterase (cG-BPDE). Specific
substrate for PKG, Km=68 µM; Km for PKA=320 µM, and no
phosphorylation detected for CaMKII or PKC122,123.
1 mg
5 x 1 mg
PKG INHIBITORS
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES)
EI-200
KT5823
98%, MW=495.5 [126643-37-6] Storage: 0°C
KT5823 is a derivative of K-252a and is a selective inhibitor of PKG (Ki=0.234
µM)193. See Table 4 for selectivity data.
100 µg
5 x 100 µg
EI-158
H-8
N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide?HCl
See "PKA inhibitors".
EI-184
HA-1004
N-(2-Guanidinoethyl)-5-isoquinolinesulfonamide?HCl
See "PKA inhibitors".
EI-233
HA-1077
1-(5-Isoquinolinesulfonyl)-homopiperazine·HCl
See "PKA inhibitors".
P-208
PKG inhibitor
Arg-Lys-Arg-Ala-Arg-Lys-Glu
>97%, MW=943.1 [82801-73-8] Storage: -20°C
Sequence is histone H2B (29-35) non-phosphorylatable analog (Ser to Ala-32) of substrate
peptide of PKG. Ki =86 µM for PKG; Ki =550 µM for PKA.
1 mg
5 x 1 mg
CaM KINASE II (CaMKII)
CaMKII ENZYME
SE-134
CaM Kinase II (Ca2+/calmodulin-dependent protein kinase II) (from rat brain)
>95% by SDS-PAGE, MW=650 kDa (EC 2.7.10.-) Storage: -70°C
Composed of 60 and 50 kDa subunits in ratios of 1:3 to 1:4. Supplied at 20 µg/ml.
Specific activity: 0.9 µmol phosphate/min/mg using synapsin I as substrate at 30°C.
Standard assay conditions are 50 mM HEPES, pH 7.4, 10 mM MgCl2, 100
µM ATP, 5 mM CaCl2, 30 µg/ml calmodulin (catalog #SE-101) and 0.6
ng/µl CaM kinase II in a total volume of 50 µl125-127.
0.5 µg
5 x 0.5 µg
CaMKII SUBSTRATES
P-101
Autocamtide-2, protein kinase substrate
Lys-Lys-Ala-Leu-Arg-Arg-Gln-Glu-Thr-Val-Asp-Ala-Leu
>97%, MW=1526.9, Storage: -20°C
Sequence derived from Ca2+/calmodulin-dependent protein kinase II
autophosphorylation site RQETVD plus basic residues to facilitate phosphocellulose
binding. Contains consensus sequence R-X-X-S/T. Highly selective CaMKII substrate (Km=2
µM for CaMKII; >50 µM for PKC; No activity towards PKA)128.
1 mg
5 x 1 mg
P-102
Syntide-2, protein kinase substrate
Pro-Leu-Ala-Arg-Thr-Leu-Ser-Val-Ala-Gly-Leu-Pro-Gly-Lys-Lys
>97%, MW=1507.8 [108334-68-5] Storage: -20°C
Phosphorylation site 2 of glycogen synthase containing the R-X-X-S/T consensus
phosphorylation site129. Substrate for CaMKII (Km
=12 µM) with purified enzyme or in cell extracts using TCA/phosphocellulose assay.
Substrate for other kinases thus inhibitors must be used to discern activity130. Reported substrate for Raf-1131-133.
1 mg
5 x 1 mg
P-100
CaM kinase IV substrate (peptide-g)
Lys-Ser-Asp-Gly-Gly-Val-Lys-Lys-Arg-Lys-Ser-Ser-Ser-Ser
>97%, MW=1450.8, Storage: -20°C
Sequence from Ca2+/calmodulin-dependent protein kinase II-g
(345-358). Substrate is specific for CaMKIV (Km=8 µM) and is a poor substrate
for type II. Useful with purified enzyme and crude extracts using phosphocellulose assay134.
1 mg
5 x 1 mg
CaMKII INHIBITORS
SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES)
EI-232
HDBA
2-Hydroxy-5-(2,5-dihydroxybenzylamino)benzoic acid
98%, MW=275.3, Storage: -20°C
Inhibits Ca2+/calmodulin kinase II (IC50=0.2 µM)135.
Also inhibits EGF receptor tyrosine kinase (IC50=0.044 µM)136
and pp60c-src(+) kinase (IC50=0.5 µM)135.
2 mg
10 mg
EI-230
KN-62
98%, MW=721.9 [127191-97-3] Storage: 0°C
KN-62 is a potent and selective inhibitor of Ca2+/calmodulin kinase II (Ki=0.9
µM). See Table 4 for selectivity data. Causes cells to arrest in S phase138.
In HEK-293 cells expressing type III adenylyl cyclase (III-AC), 10 µM blocked CaMKII
phosphorylation of III-AC139 and inhibited
hormone-stimulated Ca2+ oscillations140.
1 mg
5 x 1 mg
EI-268
KN-93
95%, MW=501.0 Storage: -20°C
KN-93 is a potent, selective and cell-permeable inhibitor of CaM kinase II (IC50
= 370 nM). Does not significantly inhibit PKA141. Causes
cell-cycle arrest in NIH 3T3 fibroblasts142 and HeLa cells143.
1 mg
EI-201
KT5926
98%, MW=525.5 [126643-38-7] Storage: 0°C
KT5926 is a propylether derivative of K-252a193. It
inhibits a number of kinases193,144 but
is most selective for CaM kinase II (Ki=4.4 nM)195.
See Table 4 for selectivity data.
100 µg
5 x 100 µg
P-200
CaM kinase II (290-309), calmodulin antagonist
Leu-Lys-Lys-Phe-Asn-Ala-Arg-Arg-Lys-Leu-Lys-Gly-Ala-Ile-Leu-Thr-Thr-Met-Leu-Ala
P-201
CaM kinase II (281-309), inhibitor
Met-His-Arg-Gln-Glu-Thr-Val-Asp-Cys-Leu-Lys-Lys-Phe-Asn-Ala-Arg-Arg-Lys-Leu-Lys-Gly-Ala-Ile-Leu-Thr-Thr-Met-Leu-Ala
>97%, MW=3374.5, Storage: -20°C
Sequence from Ca2+/calmodulin-dependent kinase A
subunit negative regulatory sequence containing the calmodulin binding site (290-309) plus
autophosphorylation site (Thr-286)145,146. Peptide can be
phosphorylated at Thr-286 by PKC147. Useful as a CaM
binding peptide and inhibits CaMKII by blocking Ca2+/CaM activation and the
enzyme active-site. IC50=80 nM, CaM binding; IC50=2 µM, active-site148.
0.5 mg
5 x 0.5 mg
P-202
CaM kinase II [Ala-286](281-302), inhibitor
Met-His-Arg-Gln-Glu-Ala-Val-Asp-Cys-Leu-Lys-Lys-Phe-Asn-Ala-Arg-Arg-Lys-Leu-Lys-Gly-Ala
>97%, MW=2600.2, Storage: -20°C
Sequence is from Ca2+/calmodulin-dependent protein kinase II
autophosphorylation site within the pseudosubstrate negative regulatory region149. Inhibition of CaMKII is as effective as 281-309 (IC50=2
µM) but kinetics are not complicated by phosphorylation of the peptide by CaMKII itself
(Thr to Ala-286)98,148.
0.5 mg
5 x 0.5 mg
P-211
AIP, CaM kinase II inhibitory peptide
>97%, MW=1498.0, Storage: -20°C
AIP (autocamtide-2-related inhibitory peptide) is a highly-specific and potent inhibitor
of CaM kinase II. This peptide is derived from the CaM kinase II substrate autocamtide-2
(BIOMOL catalog #P-101) but alanine is substituted for threonine at the 9-position. AIP at
1 µM completely inhibited CaM kinase II but did not affect PKC, PKA, CaMKIV and other
brain extract kinases150,151.
1 mg
5 x 1 mg
P-212
Myristoylated-AIP, CaM kinase II inhibitor
>97%, MW=1708.0, Storage: -20°C
This peptide is the same as AIP (P-211 above) but is N-terminal myristoylated to increase
cell-permeability. This inhibitory peptide may be highly useful in cellular studies.
0.5 mg
5 x 0.5 mg
CaMKII ANTIBODY
SA-162
Anti-Ca2+/calmodulin kinase II, a-subunit (CaMKIIa)
Mouse monoclonal antibody. Clone 6G9. Immunogen: Partially purified CaMKII152.
Supplied as purified IgG1, lyophilized. Recognizes 50 kDa a-subunit
of CaMKII in all mammalian cells plus chick and frog153,154.
Reacts with the phosphorylated and non-phosphorylated forms. Applications: WB 10 µg/ml,
IC 10 µg/ml.
100 µg
MYOSIN LIGHT CHAIN KINASE (MLCK)
MLCK SUBSTRATES
P-114
MLCK substrate, skeletal and smooth muscle
Lys-Lys-Arg-Ala-Ala-Arg-Ala-Thr-Ser-Asn-Val-Phe-Ala-NH2
>97%, MW=1418.8, Storage: -20°C
Sequence is chicken gizzard smooth muscle myosin P-light chain (MLC 11-23) with Pro to
Ala-14 and Gln to Ala-15 substitutions. Contains specificity determinants for MLC
phosphorylation (target is Ser-19) and is an excellent myosin light chain kinase (MLCK)
substrate. Km=8.6 µM for rabbit skeletal muscle MLCK, 7.5 µM for chicken
gizzard smooth muscle MLCK11,155-157.
1 mg
5 x 1 mg
P-115
MLCK substrate, skeletal muscle
Ala-Lys-Arg-Pro-Gln-Arg-Ala-Thr-Ser-Asn-Val-Phe-Ser-NH2
>97%, MW=1460.8, Storage: -20°C
Sequence is chicken gizzard smooth muscle myosin P-light chain (MLC 11-23) with Lys to
Ala-11 substitution. Myosin light chain kinase (MLCK) substrate similar to Ala-14,15
peptide but this has slightly higher Kmand lower Vmax. Km=13.3 µM
for rabbit skeletal muscle MLCK157.
1 mg
5 x 1 mg
MLCK INHIBITORS
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES)
EI-197
ML-7
1-(5-Iodonaphthalene-1-sulfonyl)-1H-hexahydro-l,4-diazepine·HCl
99%, MW=452.7 [109376-83-2] Storage: 0°C
Potent and selective inhibitor of MLC kinase (Ki=0.3 µM)158.
See Table 4 for selectivity data.
10 mg
50 mg
EI-153
ML-9
1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-l,4-diazepine·HCl
99%, MW=361.3 [105637-50-1] Storage: 0°C
Selective inhibitor of MLC kinase (Ki=3.8 µM)158,159.
Inhibits insulin-stimulated 2-deoxyglucose transport160 and
PP-1 activation161 in adipocytes, possibly via an effect on
MAP kinase or its activation160. See Table 4 for
selectivity data.
10 mg
50 mg
P-209
MLCK inhibitor
Lys-Arg-Arg-Trp-Lys-Lys-Asn-Phe-Ile-Ala-Val-NH2
>97%, MW=1444.9, Storage: -20°C
Sequence from skeletal muscle myosin light chain kinase (MLCK) (342-352) pseudosubstrate
site. Inhibits MLCK (Ki =1 µM with MLC peptide substrate). Possesses only
slight calmodulin binding activity162,155.
1 mg
5 x 1 mg
CASEIN KINASE II ENZYME AND REAGENTS
SE-124
Casein kinase II (human, recombinant)
MW=44 and 26 kDa subunits (EC 2.7.1.37) Storage: -70°C
Expressed in E. coli. Supplied in Tris/NaCl/0.1% Triton X-100. Consists of 44 kDa a catalytic and 26 kDa b regulatory
subunits as an a2b2
tetramer169. Free from proteases and phosphatases. Supplied
at 0.7 mg/ml. Specific activity: 700 kU/mg protein. One unit will transfer 1 pmol
phosphate from ATP to peptide substrate per minute at 30°C163.
10 kU
5 x 10 kU
P-113
Casein kinase II peptide substrate
Arg-Arg-Glu-Glu-Glu-Thr-Glu-Glu-Glu
>97%, MW=1206.3, Storage: -20°C
Sequenced based on CKII phosphorylation sites in casein. Substrate is specific for CKII (Km=0.5
mM), not PKA, PKG, phosphorylase kinase, MLCK, RPTK, or CKI164.
Used in phosphocellulose assay with crude extracts of bovine liver165.
1 mg
5 x 1 mg
EI-231
DRB
5,6-Dichloro-1-b-D-ribofuranosylbenzimidazole
98%, MW=319.1 [53-85-0] Storage: -20°C
DRB is a potent and specific inhibitor of casein kinase II (CKII, IC50~6 µM).
It has been used to inhibit RNA polymerase II transcription which may be dependent on CKII166 and its interaction with ATF-1167.
10 mg
50 mg
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES
SA-163
Anti-Casein kinase II, a-subunit (CKIIa)
Mouse monoclonal antibody. Clone 1AD9. Immunogen: Recombinant CKIIa168,169. Supplied as purified IgG1, lyophilized. Recognizes
p40-44 kDa catalytic a-subunit of CKII. Applications: WB 1-5
µg/ml.
100 µg
MAP KINASE (MAPK)
MAPK ENZYME
SE-137
MAP Kinase, activated (rat, recombinant)
>95% by SDS-PAGE, MW=42 kDa, Storage: -70°C
This is the ERK2 (p42mapk) isoform cloned from a rat skeletal muscle cDNA
library. It is constructed with a 6-His N-terminal tag to facilitate purification, and
expressed in E. coli. The enzyme was then activated by phosphorylation in vitro
using a constitutively active MEK1 mutant. The MAP kinase was repurified and contains no
contaminating MEK1 as detected by SDS-PAGE and Western blotting. Supplied at 0.1 mg/ml.
Specific activity: ~2 µmol phosphate/min/mg at 30°C using myelin basic protein (MBP) as
substrate. Useful to study enzyme regulation, kinetics, and phosphorylate target
substrates170-172.
5 µg
5 x 5 µg
SE-139
MAP Kinase, nonactivated (rat, recombinant)
>95% by SDS-PAGE, MW=42 kDa, Storage: -70°C
Identical to SE-137 above, but not activated by MEK1. Supplied at 0.5 mg/ml. Specific
activity: ~2 nmol phosphate/min/mg at 30°C using myelin basic protein (MBP) as substrate.
Potential activation by MEK1 is 1000-fold. Useful to study enzyme activation by other
kinases and as a control for activated MAP kinase (SE-137).
20 µg
5 x 20 µg
MAPK SUBSTRATES
P-121
EGF-R (661-681) T669 Peptide, MAP Kinase Substrate
Lys-Arg-Glu-Leu-Val-Glu-Pro-Leu-Thr-Pro-Ser-Gly-Glu-Ala-Pro-Asn-Gln-Ala-Leu-Leu-Arg
>97%, MW=2315.0 Storage: -20°C
Peptide sequence is based on the cytoplasmic domain of the EGF-receptor, inclusive of
Thr-669, a major phosphorylation site in cells stimulated by IL-1, PMA and EGF173,174. This is an excellent substrate for MAP kinase but a
poor substrate for MAPKK175.
1 mg
5 x 1 mg
P-122
MBP (95-98) S5 Peptide, MAP Kinase Substrate
Ala-Pro-Arg-Thr-Pro-Gly-Gly-Arg-Arg
>97%, MW=967.0, Storage: -20°C
Peptide sequence is based on myelin basic protein (MBP) residues 95-98 inclusive of
Thr-97, the major site of phosphorylation by MAP kinase in vitro176-178.
1 mg
5 x 1 mg
P-124
Erk1/Erk2 Peptide, MAP kinase kinase substrate
Ala-Asp-Pro-Asp-His-Asp-His-Thr-Gly-Phe-Leu-Thr-Glu-Tyr-Val-Ala-Thr-Arg-Trp-Arg-Arg
>97%, MW=2543.8, Storage: -20°C
Sequence is based on Erk1/Erk2 residues 172-190 MAP kinase kinase phosphorylation site
(underlined)179. N-terminal Arg is added for
phosphocellulose binding180.
1 mg
5 x 1 mg
MAPK ANTISENSE OLIGO'S
ZA-100
MAP Kinase Antisense Oligo
>90% by HPLC and capillary electrophoresis, MW=5.2 kDa, Storage: -70°C.
This is a 17-mer phosphorothioate oligodeoxynucleotide. It is directed against a sequence
which is identical in the p42 and p44 MAP kinase isoforms (ERK2, ERK1) and conserved in
human, mouse and rat. When complexed with the cationic lipid DOTMA and applied at 2 µM,
it eliminated immunodetectable p42 and p44 MAP kinases from 3T3 L1 fibroblasts in 48 hrs.181. Differentiation of the fibroblasts into adipocytes was
blocked at > 1 µM181. It depleted MAP kinase >90%
in rat aortic smooth muscle cells (DOTAP complex, 5-10 µM)182.
A 10 µM treatment of these cells eliminated angiotensin II or PDGF-stimulated MAP kinase
activity and reduced PDGF-stimulated DNA synthesis by 95%. There was no effect on
expression of the MAP kinase homologs, p38 and JNK, and no effect on MEK activation182. Supplied lyophilized.
25 nmols
ZA-101
MAP Kinase Control Oligo
>90% by HPLC and capillary electrophoresis, MW=5.2 kDa, Storage: -70°C.
Phosphorothioate 17-mer oligodeoxynucleotide with the same base composition as the MAP
kinase antisense oligonucleotide ZA-100 above, but is a randomized sequence. Does not
affect MAP kinase expression or activity181,182. Supplied
lyophilized.
25 nmols
ZP-100
MAP Kinase Antisense and Control Oligo Pair
Twenty-five nanomoles each of ZA-100 and ZA-101, above.
1 pair
MAPK INHIBITORS
(SEE TABLE 4 FOR SER/THR KINASE INHIBITOR SPECIFICITIES)
EI-360
PD-98059
>95%, MW=267.3 [167869-21-8] Storage: -20?/font>C
PD-98059 is a new, potent and selective inhibitor of MAP kinase kinase (MEK). It
selectively blocks the activation of MEK thereby inhibiting the phosphorylation and the
activation of MAP kinase183,184. PD-98059 is cell
permeable. In PC12 pheochromocytoma cells, it completely blocked the increase in MAP
kinase activity produced by NGF, IC50=2 mM183,185. PD-98059 is an invaluable, new tool to help elucidate
the role of the MAPK cascade in a variety of biological systems.
5 mg
5 x 5 mg
JUN KINASE
SE-150
C-JUN N-terminal kinase (rat, recombinant)
>95% by SDS-PAGE, MW=52 kDa, Storage: -70°C
This c-jun kinase (JNK3, SAPK-b, p54b)
was cloned from a rat skeletal muscle cDNA library and expressed in E. coli186. The expression vector encoded a fusion protein comprising
(from N- to C-terminus): A calmodulin binding peptide (rat MLCK residues 579-605),
thrombin cleavage site, and JNK3 (MW=48.1 kDa). The recombinant protein was purified by
affinity chromatography on a calmodulin resin. Supplied at 0.4 mg/ml. Specific activity:
40 nmol phosphate/min/mg at 30°C using GST-c-jun (1-79) fragment (catalog #SE-151, below)
as substrate. Useful to study enzyme regulation, kinetics, and phosphorylate target
substrates.
20 µg
5 x 20 µg
SE-151
C-JUN (1-79), c-jun kinase substrate (rat, recombinant)
>95% by SDS-PAGE, MW=37 kDa, Storage: -70°C
This is a GST-fusion protein of the c-jun (1-79) activation domain, expressed in E.
coli. It is a highly specific substrate for JNK/SAPK It is not appreciably
phosphorylated by MAP kinase or by p38 kinase. It can be used as a substrate for c-jun
kinase (catalog SE-150, above) at 2 µg per assay187,188.
Supplied at 1 mg/ml.
250 µg
5 x 250 µg
SER/THR KINASES - BROAD SPECIFICITY INHIBITORS
EI-145
A-3
N-(2-Aminoethyl)-5-chloronaphthalene-1-sulfonamide·HCl
98%, MW=321.23, Storage: 0°C
Inhibits PKA (Ki=4.3 µM), PKG (Ki= 3.8 µM), MLCK (Ki=7.4
µM) and PKC (Ki=47 µM)189. See Table 4.
10 mg
50 mg
EI-148
H-7
1-(5-Isoquinolinesulfonyl)-2-methylpiperazine?HCl
99%, MW=364.3 [84477-87-2] Storage: 0°C
Inhibits PKA (Ki=3.0 µM), PKG (Ki=5.8 µM), MLCK (Ki=97
µM) and PKC (Ki=6 µM)190. See Table 4.
10 mg
50 mg
EI-195
H-9
N-(2-Aminoethyl)-5-isoquinolinesulfonamide?HCl
99%, MW=324.3 [84468-17-7] Storage: 0°C
H-9 is an excellent affinity ligand for chromatographic purification of protein kinases191,192. It also inhibits protein kinases190.
See Table 4 for selectivity data.
10 mg
50 mg
EI-152
K-252a
99%, MW=467.5 [97161-97-2] Storage: 0°C
Inhibits PKA (Ki=0.018 µM), PKG (Ki=0.020 µM), MLCK (Ki=0.017
µM), PKC (Ki=0.025 µM)193,194, CaMK (Ki=0.0018
µM)195 and phosphorylase kinase (IC50=1.7 nM)196. See Table 4. Potent inhibitor of the trk family of
receptor tyrosine kinases197,198,199 and MAP kinase200.
100 µg
1 mg
EI-234
K-252b
99%, MW=453.5 [99570-78-2] Storage: 0°C
Inhibits PKA (Ki=0.09 µM), PKG (Ki=0.1 µM), MLCK (Ki=0.147
µM) and PKC (Ki=0.02 µM)193. See Table 4.
100 µg
1 mg
EI-156
Staurosporin
99%, MW=466.5 [62996-74-1] Storage: 0°C
Staurosporin inhibits a variety of kinases including PKA (Ki=7.0 nM), PKG (Ki=8.5
nM), MLCK (Ki=1.3 nM), PKC (Ki=0.7 nM)201,202,
CaMK (IC50=20 nM)203, tyrosine kinases (IC50=70
nM)204,205 and phosphorylase kinase (IC50=0.5
nM)196. See Table 4. Inhibition is via interaction with the
ATP binding site206. It induces PKC translocation207 and augments PMA induced ornithine decarboxylase208. Activates a bcl-2-regulated apoptosis pathway209.
100 µg
1 mg
NEW LOWER PRICE
TABLE 4. SPECIFICITIES OF SELECTED SER/THR KINASE INHIBITORS.
[Inhibition as Ki or (IC50), concentration in µM]
| Kinase |
PKA |
PKC |
PKG |
MLCK |
CaMKII |
CK-I |
CK-II |
| Inhibitor |
|||||||
| A-3 |
4.3 |
47 |
3.8 |
7.4 |
80 |
5.1 |
|
| Calphostin C |
(>50) |
(0.05) |
(>25) |
(>5) |
|||
| Chelerythrine |
(170) |
(0.66) |
(>100) |
||||
| GF 109203X |
(2) |
(0.02) |
|||||
| H-7 |
3.0 |
6.0 |
5.8 |
97 |
|||
| H-8 |
1.2 |
15 |
0.48 |
68 |
|||
| H-9 |
1.9 |
18 |
0.87 |
70 |
|||
| H-89 |
0.048 |
31.7 |
0.48 |
28.3 |
29.7 |
38.3 |
137 |
| HA-1004 |
2.3 |
40 |
1.3 |
150 |
|||
| HA-1077 |
1.6 |
1.6 |
36 |
||||
| Hypericin |
(>79) |
(3.4) |
(>40) |
||||
| K-252a |
0.018 |
0.025 |
0.020 |
0.017 |
0.0018 |
||
| K- 252b |
0.09 |
0.02 |
0.1 |
0.147 |
|||
| KN-62 |
>100 |
>100 |
>100 |
0.9 |
|||
| KT5720 |
0.056 |
>2 |
>2 |
>2 |
|||
| KT5823 |
>10 |
4 |
0.234 |
>10 |
|||
| KT5926 |
1.2 |
0.723 |
0.158 |
0.018 |
0.0044 |
||
| ML-7 |
21 |
42 |
0.3 |
||||
| ML-9 |
32 |
54 |
3.8 |
||||
| Staurosporin |
0.007 |
0.0007 |
0.0085 |
0.0013 |
(0.02) |
Abbreviations used are: PKA, cAMP-dependent protein kinase; PKC, protein kinase C; PKG,
cGMP-dependent protein kinase; MLCK, myosin light-chain kinase; CaMKII, Ca2+/calmodulin
kinase II; CK-I, casein kinase I; CK-II, casein kinase II.
SER/THR KINASES - MISCELLANEOUS REAGENTS
P-116
S6 peptide, protein kinase substrate
Arg-Arg-Leu-Ser-Ser-Leu-Arg-Ala
>97%, MW=958.2, Storage: -20°C
Sequence is from rat hepatic 40S ribosomal protein S6 (232-239). Substrate for p90 rsk
(S6K) in cell extracts (Km=0.18 mM) using phosphocellulose assay. Peptide
phosphorylation by PKA and PKC can be eliminated by the inclusion of PKI and EGTA,
respectively, in the assay210-212.
1 mg
5 x 1 mg
ST-101
Adenosine-5'-O-(3-thiotriphosphate)·Li4 (ATPg-S)
>85%, MW=547.0 [93839-89-5] Storage: -20°C
An ATP analog that can substitute for ATP in various kinase reactions. The resulting
thiophosphorylated proteins are resistant to dephosphorylation by protein phosphatases213.
5 mg
25 mg
ST-102
Anisomycin
97%, MW=265.3 [22862-76-6] Storage: -20°C
Activates JNK/SAPKs but not ERKs leading to the phosphorylation of S6, histone H3 and
HMG-14 and the induction of the immediate-early genes c-fos and c-jun214-216.
10 mg
50 mg
EI-144
Mezerein
99%, MW=654.7 [34807-41-5] Storage: -20°C
A non-phorbol ester diterpene comparable to PMA as an inflammatory agent but less potent
as a tumor promotor217. A potent "second stage"
tumor promotor218.
1 mg
5 mg
TYROSINE KINASES
A great deal of attention is currently being focused on the role of tyrosine kinases219 in receptor-mediated signal transduction220,221.
Several oncogene products219,222 as well as a number of
receptors for growth factors including EGF223, PDGF224, NGF225 and insulin226 are tyrosine kinases. In platelets, both PAF227 and a thromboxane A2 agonist228
were found to stimulate tyrosine phosphorylation as were thrombin229
and collagen230. In human neutrophils, the chemotactic
factor fMLP and GM-CSF both induced tyrosine phosphorylation of several proteins231. In lymphocytes including T cells232,233,
B cells234 and NK cells235,
tyrosine phosphorylation was shown to be an early, critical event in receptor mediated
activation.
EPIDERMAL GROWTH FACTOR RECEPTOR (EGFR)
EGFR ENZYME
SE-167
EGF Receptor Kinase Domain, protein tyrosine kinase (human, recombinant)
Inquire
SE-116
EGF Receptor, protein tyrosine kinase (from human A431 cells)
>90% by SDS-PAGE, MW=170 kDa, Storage: -70°C
Supplied in 10% glycerol, 0.05% Triton X-100 buffer containing 5 mM EGF. EGFR is active
but not autophosphorylated. Specific activity: 45 U/ml. One unit will add one pmol
phosphate to angiotensin II per minute at pH 7.4 and 30°C.
10 U
EGFR INHIBITORS
(SEE TABLE 5 FOR TYROSINE KINASE INHIBITOR SPECIFICITIES)
EI-228
AG-494
99%, MW=280.3, Storage: RT
AG-494 is a member of the tyrphostin family of tyrosine kinase inhibitors and is a potent
inhibitor of EGF receptor autophosphorylation (IC50=1.2 µM) and EGF-dependent
cell growth (IC50=6 µM)236. It selectively
inhibits HER1 (EGF receptor) vs. HER1-2 receptor autophosphorylation. HER1: IC50=1.1
µM; HER1-2: IC50=45 µM237. HER1-2 is a
chimeric receptor consisting of the external HER1 domain fused to an internal HER2 domain.
See Table 5 for selectivity data.
10 mg
50 mg
EI-267
AG-825
5-[(Benzthiazol-2-yl)thiomethyl]-4-hydroxy-3-methoxybenzylidenecyanoacetamide
98%, MW=397.5, Storage: RT
A selective HER1-2 (HER2-Neu)238 tyrosine kinase inhibitor237. See Table 5 for selectivity data.
2 mg
10 mg
EI-277
AG-1478
4-(3-Chloroanilino)-6,7-dimethoxyquinazoline
>98%, MW=315.8 [63177-57-1] Storage: -20°C
Potent and selective inhibitor of EGFR (IC50=3 nM)238.
It reduces EGF-stimulated DNA synthesis in Rat-1 fibroblasts by ~75% at 0.25 µM239. Blocks EGF-dependent src-family kinase activation240 and p21/Cip1/WAF1 induction241
in A431 cells. Also inhibits H2O2 -induced stimulation of
amphiregulin expression in rat gastric epithelium (RGM 1) cells242.
See Table 5 for selectivity data.
5 mg
25 mg
EI-146
Erbstatin Analog
Methyl 2,5-dihydroxycinnamate
98%, MW=194.0 [63177-57-1] Storage: RT
Erbstatin is a novel inhibitor of the EGF receptor associated tyrosine kinase243. It is an unstable compound and is completely inactivated
in serum in 30 minutes. Methyl 2,5-dihydroxycinnamate is a stable erbstatin analog
retaining activity after a 60 minute incubation. It inhibits EGF receptor associated
tyrosine kinase in vitro (IC50=0.77 µM)244.
Inhibition is competitive with substrate and noncompetitive with ATP.
5 mg
25 mg
EI-232
HDBA
2-Hydroxy-5-(2,5-dihydroxybenzylamino)benzoic acid
Inhibits EGF receptor tyrosine kinase (IC50=0.044 µM)136
and pp60c-src(+) kinase (IC50=0.5 µM)135.
Also inhibits Ca2+/calmodulin kinase II (IC50=0.2 µM)135.
EI-185
Lavendustin A
98%, MW=381.4 [125697-92-9] Storage: 0°C
Lavendustin A is a potent and selective inhibitor of the EGF receptor tyrosine kinase (IC50=11
nM) It binds to a site on the kinase which is distinct from the ATPand peptide substrate binding sites246. Inhibited EGF-induced proliferation of cultured human
myometrial smooth muscle cells247. Suppressed VEGF-induced
angiogenesis in rats248. See Table 5 for selectivity data.
1 mg
5 mg
EI-252
RG-13022
99%, MW=266.3 [136831-48-6] Storage: RT
RG-13022 is a non-phenolic tyrphostin analog which inhibits the EGF receptor (IC50=5
µM) and PDGF receptor tyrosine kinases and is long acting. It inhibits EGF-stimulated
HER14 cell proliferation (IC50=1 µM) as well as tumor growth in vivo249. It inhibits both EGF- and PDGF-stimulated DNA synthesis
in human glioma cell lines. RG-13022 is non-phenolic and therefore possesses less
antioxidant activity than other tyrphostins.
5 mg
25 mg
EI-253
RG-14620
99%, MW=274.1 [136831-49-7] Storage: RT
RG-14620 is a non-phenolic tyrphostin analog which is selective for the EGF receptor and
long acting. It inhibits EGF-stimulated HER14 cell proliferation (IC50=3 µM)
as well as tumor growth in vivo249. Has
anti-proliferative effects on renal and bladder carcinoma cell lines251.
5 mg
25 mg
EI-191
Tyrphostin 23 (RG-50810)
99%, MW=186.1 [118409-57-7] Storage: RT
A potent inhibitor of EGF receptor kinase activity252,253,257.
See Table 5 for selectivity data.
10 mg
50 mg
EI-187
Tyrphostin 25 (RG-50875)
99%, MW=202.1 [118409-58-8] Storage: RT
A potent inhibitor of EGF receptor kinase activity253,257,296. See Table 5 for selectivity data.
5 mg
25 mg
EI-257
Tyrphostin 46
99%, MW=204.2 [122520-85-8] Storage: RT
Inhibits EGF receptor kinase286, p56lck254
and PDGF255 receptor kinase. See Table 5 for data.
10 mg
50 mg
EI-188
Tyrphostin 47 (RG-50864, AG-213)
99%, MW=220.2 [122520-86-9] Storage: RT
A potent inhibitor of EGF receptor kinase activity252,253,257.
See Table 5 for selectivity data.
5 mg
25 mg
EI-189
Tyrphostin 51
99%, MW=268.2 [122520-90-5] Storage: 0°C
A potent inhibitor of EGF receptor kinase activity. Inhibition is mixed competitive with
ATP and substrate253,256,257. See Table 5 for selectivity
data.
5 mg
25 mg
EI-190
Tyrphostin 1
99%, MW=184.2 [2826-26-8] Storage: RT
An inactive tyrphostin which may be used as a negative control257.
See Table 5.
20 mg
100 mg
EGFR ANTIBODIES
SA-110
Anti-Epidermal growth factor receptor (EGFR)
Mouse monoclonal antibody. Clone EGFR1. Immunogen: Whole A431 human epidermal carcinoma
cells258. Supplied as purified IgG2b, 0.1 mg/ml. Reacts
with human EGFR 175 kDa extracellular domain. Does not inhibit EGF binding. Applications:
WB, IP, IF.
200 µg
SA-111
Anti-Epidermal growth factor receptor (EGFR)
Mouse monoclonal antibody. Clone F4. Immunogen: Human EGFR (985-996) peptide259. Supplied as purified IgG1, 0.1 mg/ml. Reacts with 170 kDa
EGFR intracellular domain. Recognizes autophosphorylated EGFR and the related v-erb
B protein, but the immunoprecipitated EGFR can not autophosphorylate. Applications: IP,
IF, IH, ELISA.
200 µg
INSULIN RECEPTOR
The insulin receptor is a tyrosine kinase that is present in virtually all vertebrate
tissues. It is composed of two a-subunits that are each linked
to a b-subunit and to each other by disulfide bonds. The a-subunits are located extracellularly and contain the insulin
binding site, whereas the intracellular portion of the b-subunit
contains the protein tyrosine kinase domain. Regulatory autophosphoryation sites in the b-subunit consists of Tyr 1146, 1150, and 1151. Autophosphorylation
of all 3 tyrosine residues in the YXXXYY motif of this region stimulates kinase activity
10- to 20-fold260. The following phosphopeptides correspond
to the insulin receptor regulatory region. These peptides can be used to study insulin
receptor signalling. In addition, they have been used as tyrosine phosphatase substrates
in non-radioactive PTPase assays261.
INSULIN RECEPTOR ENZYME
SE-159
b-Insulin Receptor Kinase (b-IRK),
protein tyrosine kinase (human, recombinant)
48 kDa cytoplasmic domain of the b-subunit (resudues 941-1343),
expressed in a baculovirus expression system.
Inquire
INSULIN RECEPTOR SUBSTRATES
P-314
IR0, Insulin Receptor [1142-1153]
Thr-Arg-Asp-Ile-Tyr-Glu-Thr-Asp-Tyr-Tyr-Arg-Lys
>97%, MW=1622.9, Storage: -20°C
Sequence is from insulin receptor b-subunit cytoplasmic domain,
inclusive of regulatory autophosphorylation sites Tyr 1146, 1150 and 1151260.
It has been used as an insulin receptor tyrosine kinase substrate262,261
and the phosphorylated peptide as a protein tyrosine phosphatase substrate.
0.5 mg
5 x 0.5 mg
P-315
IR5, Insulin Receptor [1142-1153] (pY1146)
Thr-Arg-Asp-Ile-pTyr-Glu-Thr-Asp-Tyr-Tyr-Arg-Lys
>97%, MW=1703.0, Storage: -20°C
0.5 mg
5 x 0.5 mg
P-316
IR9, Insulin Receptor [1142-1153] (pY1150)
Thr-Arg-Asp-Ile-Tyr-Glu-Thr-Asp-pTyr-Tyr-Arg-Lys
>97%, MW=1703.0, Storage: -20°C
0.5 mg
5 x 0.5 mg
P-317
IR10, Insulin Receptor [1142-1153] (pY1151)
Thr-Arg-Asp-Ile-Tyr-Glu-Thr-Asp-Tyr-pTyr-Arg-Lys
>97%, MW=1703.0, Storage: -20°C
0.5 mg
5 x 0.5 mg
P-318
IR5,9,10, Insulin Receptor [1142-1153] (pY1146,1150,1151)
Thr-Arg-Asp-Ile-pTyr-Glu-Thr-Asp-pTyr-pTyr-Arg-Lys
>97%, MW=1863.0, Storage: -20°C
0.5 mg
5 x 0.5 mg
P-319
Insulin Receptor Phosphopeptide Kit
Contains 0.5 mg each of the above peptides: (P-314, P-315, P-316, P-317, P-318)
1 kit
P-320
IRS-1 (Y608) peptide
Lys-Lys-His-Thr-Asp-Asp-Gly-Tyr-Met-Pro-Met-Ser-Pro-Gly-Val-Ala
>97%, MW=1732.8, Storage: -20°C
Sequence is from insulin receptor substrate 1 (IRS-1) inclusive of Tyr-608. It contains
the insulin receptor tyrosine kinase substrate motif YMXM (Tyr-Met-X-Met). This peptide
has been used as a substrate for purified insulin receptor (Km=90 µM)263 and other tyrosine kinases264
in phosphocellulose binding assays. The tyrosine phosphorylated version of this peptide
binds to phosphatidylinositol 3-kinase (PI 3-kinase) SH2 domain and activates the enzyme265.
1 mg
5 x 1 mg
INSULIN RECEPTOR INHIBITORS
(SEE TABLE 5 FOR TYROSINE KINASE INHIBITOR SPECIFICITIES)
EI-247
HNMPA
Hydroxy-2-naphthalenylmethylphosphonic acid
98%, MW=238.2, Storage: -20°C
HNMPA is a membrane impermeable inhibitor of insulin receptor tyrosine kinase activity (IC50=100
µM)266. It was recently shown to inhibit both tyrosine and
serine autophosphorylation by the human insulin receptor267.
5 mg
25 mg
EI-248
HNMPA-(AM)3
Hydroxy-2-naphthalenylmethylphosphonic acid-tris acetoxymethylester
98%, MW=454.4, Storage: -20°C
HNMPA-(AM)3 is a cell permeable analog of HNMPA. It inhibits insulin receptor
tyrosine kinase activity (IC50=100 µM) and insulin-stimulated glucose
oxidation in isolated rat adipocytes (IC50=10 µM). It has no effect on PKA (at
concs. up to 1 mM) or PKC (at concs. up to 420 µM)266.
5 mg
25 mg
SRC-FAMILY
SRC-FAMILY INHIBITORS
(SEE TABLE 5 FOR TYROSINE KINASE INHIBITOR SPECIFICITIES)
EI-274
Damnacanthal
95%, MW=282.3 [477-84-9] Storage: -20?/font>C
Damnacanthal is the most potent and selective inhibitor of p56lck tyrosine
kinase described to date268. It inhibits p56lck
autophosphorylation (IC50=17 nM) as well as phosphorylation of exogenous
substrates (IC50=620 nM) in cell-free assays. Damnacanthal shows a >10-fold
selectivity for p56lck over PKA and PKC and >40-fold selectivity over four
receptor tyrosine kinases as well as 7-20-fold selectivity over the homologous enzymes p60src
and p59fyn. Damnacanthol is not active in whole cell tyrosine kinase assays.
1 mg
5 x 1 mg
EI-227
Herbimycin A
98%, MW=574.3 [70563-58-5] Storage: -20°C
Herbimycin A, a benzoquinoid ansamycin antibiotic, irreversibly and selectively inhibits
tyrosine kinases by reacting with thiol groups269,270. It
is effective on Src, Yes, Fps, Ros, Abl and ErbB oncogene products269,270,271
and inhibits thrombin-induced tyrosine phosphorylation of phospholipase C272.
100 µg
1 mg
EI-271
Piceatannol
3,4,3',5'-Tetrahydroxy-trans-stilbene
98%, MW=244.3 [10083-24-6] Storage -20?/font>C
Piceatannol preferentially inhibits the activity of Syk (IC50~10 mM) as compared with Lyn in isolated enzyme preparations. In RBL-2H3
cells the selective inhibition of Syk by piceatannol results in inhibition of FceR1-mediated signaling273.
1 mg
5 x 1 mg
EI-275
PP1
98%, MW=281.4, Storage: -20?/font>C
A highly potent and selective inhibitor of Src family tyrosine kinases, IC50=5
nM for p56lck; p59lyn=6 nM; p60src=170 nM. Inhibition of
T-cell phosphorylation in response to anti-CD3 activation occurs at 10 µM274.
1 mg
5 mg
SRC-FAMILY PEPTIDES
P-307
pp60v-src autophosphorylation site, protein tyrosine kinase substrate
Arg-Arg-Leu-Ile-Glu-Asp-Asn-Glu-Tyr-Thr-Ala-Arg-Gly
>97%, MW=1592.9 [81493-98-3] Storage: -20°C
Residues 2-12 are the v-src (412-422) autophosphorylation site with an extra Arg
for phosphocellulose binding. This substrate was originally described as an EGF-receptor
substrate in A431 cell extracts (Km=0.3-0.6 mM)275.
1 mg
5 x 1 mg
P-312
pp60v-src autophosphorylation site, tyrosine phosphopeptide
Arg-Arg-Leu-Ile-Glu-Asp-Asn-Glu-Tyr(PO3H2)-Thr-Ala-Arg-Gly
>97%, MW=1671.9, Storage: -20°C
Same as P-307 but this peptide is tyrosine phosphorylated. Useful in a non-radiolabeled
tyrosine phosphatase assay and as a competitive tyrosine phosphatase inhibitor.
1 mg
5 x 1 mg
P-308
RR-SRC, protein tyrosine kinase substrate
Arg-Arg-Leu-Ile-Glu-Asp-Ala-Glu-Tyr-Ala-Ala-Arg-Gly
>97%, MW=1519.9 [81156-93-6] Storage: -20°C
Tyrosine kinase substrate derived from v-src autophosphorylation site (original src
sequence is IEDNEYTARQG). Km=5 mM in LSTRA cell extracts276.
1 mg
5 x 1 mg
P-306
Peptide A, protein tyrosine kinase inhibitor
Val-Ala-Pro-Ser-Asp-Ser-Ile-Gln-Ala-Glu-Glu-Trp-Tyr-Phe-Gly-Lys-Ile-Thr-Arg-Arg-Glu
>97%, MW=2482.8, Storage: -20°C
Sequence is pp60v-src (137-157) which is located in the SH3/SH2 domain
region277. Non-competitive inhibitor of src and
EGF-R278. Selectively inhibits protein tyrosine
phosphorylation in cell extracts. Peptide is not phosphorylated by src. IC50=7.5
µM for v-src. Inhibitory target seems to be a catalytic negative regulatory domain
of the tyrosine kinase279.
0.5 mg
5 x 0.5 mg
SRC-FAMILY ANTIBODIES
SA-137
Anti-Src, human endogenous
Rabbit polyclonal antibody. Immunogen: Rous sarcoma virus (RSV). Supplied as serum.
Specificity: pp60c-src from human, rat, mouse and chick plus other viral
proteins of RSV. Can detect src in fibroblasts, endothelial and neuronal cells.
Applications: WB 1/1000, IF 1/100, IP.
200 µl
SA-248
Anti-Syk
Mouse monoclonal antibody. Clone 4D10.1. Immunogen: Human Syk (313-339) peptide conjugated
to KLH. Supplied as purified IgG2a, 0.2 mg/ml. Recognizes 72 kDa human syk tyrosine
kinase. Does not crossreact with pig or mouse. Applications: WB 1 µg/ml, IP (active in
immune-complex kinase assays), IH.
100 µg
OTHER TYROSINE KINASE INHIBITORS
EI-272
AG-490
98%, MW=294.3 Storage: -20?/font>C
AG-490 is a potent inhibitor of the JAK-2 tyrosine kinase. In acute lymphoblastic leukemia
(ALL) cells which abundantly express JAK-2, AG-490 dose-dependently inhibited DNA
synthesis, blocked cell growth and induced apoptosis. At 5 mM,
AG-490 almost completely blocked the growth of all pre-B ALL cells but had no significant
effect on the growth of mitogen-stimulated normal B or T cells, B-cell lymphoma or T-cell
leukemia cells. AG-490 does not significantly inhibit other kinases such as Lck, Lyn, Btk,
Syk and Src280. It blocks interleukin-7-induced JAK kinase
activity in T-cells (JAK-1, JAK-3) and the consequent phosphorylation of PI-3 kinase281,282. AG-490 is cell permeable and is a valuable new tool
for studying the cellular role of JAK kinases in signal transduction.
5 mg
25 mg
EI-263
AG-126
98%, MW=215.2 Storage: RT
AG-126 prevents lipopolysaccharide-induced lethal toxicity (septic shock) in mice.
Administered at 1.5 mg/mouse, it reduced LPS-induced lethal toxicity from 95% to 10%.
AG-126 greatly reduces LPS-induced tyrosine phosphorylation of p42MAPK in
murine peritoneal macrophages at 50 µM. EGFR, Her-2/neu receptor or PDGF-R are not
inhibited at 100 µM283. Inhibits tyrosine kinase-dependent
B cell receptor signalling284,285.
5 mg
25 mg
EI-229
AG-370
99%, MW=259.3 Storage: 0°C
AG-370 is a member of the tyrphostin family of tyrosine kinase inhibitors and is a
selective inhibitor of the PDGF receptor kinase (IC50=20 µM) vs. the EGF
receptor kinase286. AG-370 inhibits PDGF-induced
mitogenesis in human bone marrow fibroblasts287. See Table
5 for selectivity data.
2 mg
10 mg
EI-258
AG-879
99%, MW=316.5, Storage: -20°C
AG-879, a member of the tyrphostin family of tyrosine kinase inhibitors, inhibits NGF
receptor (pp140c-trk) autophosphorylation selectively with no inhibition
of EGF or PDGF receptor phosphorylation (IC50=10 µM)288.
AG-879 inhibits NGF-induced neurite outgrowth in PC12 cells288.
See Table 5 for selectivity data.
5 mg
25 mg
EI-147
Genistein
4',5,7-Trihydroxyisoflavone
98%, MW=270.2 [446-72-0] Storage: RT
Genistein inhibits tyrosine phosphorylation in isolated enzyme and receptor preparations
and in whole cells including platelets227,228, lymphocytes232,233,234 and a variety of cultured cells289-292.
In cultured A431 epidermoid carcinoma cells, EGF stimulated tyrosine phosphorylation was
totally inhibited by genistein at 100 µg/ml292. Inhibition
is competitive with ATP and noncompetitive with substrate (see Table 5 for selectivity).
20 mg
100 mg
ST-110
Daidzein
4',7-Dihydroxyisoflavone
98%, MW=254.2 [486-66-8] Storage: RT
Inactive negative control compound for genistein292. Shares
with genistein, a number of effects thought to be unrelated to tyrosine kinase inhibition.
These include activation of the human estrogen receptor293,
induction of differentiation in leukemia cells294 and
inhibition of fast Na+ channels295.
20 mg
100 mg
EI-215
Tyrphostin 9 (RG-50872, Malonaben, SF 6847)
99%, MW=282.4 [10537-47-0] Storage: RT
Tyrphostin 9 is a selective inhibitor of the PDGF receptor tyrosine kinase (IC50=1.2
µM)286,296. It is also a potent (10
nM) uncoupler of oxidative phosphorylation297. See Table 5
for selectivity data.
50 mg
250 mg
TABLE 5. SELECTIVITY OF TYROSINE PROTEIN KINASE INHIBITORS
| Catalog # |
Compound |
Kinase |
IC50(µM) |
Substrate |
Ref. |
| EI-229 |
AG-370 |
EGFR |
820 |
poly-GAT |
286 |
| PDGFR |
20 |
unspecified |
286 |
||
| EI-228 |
AG-494 |
EGFR |
0.7 1.2 |
poly-GAT autophos. |
236 |
| HER1-2 |
42 |
autophos. |
298 | ||
| PDGFR |
6 |
autophos. |
237 |
||
| IR |
>100 |
unspecified |
237 |
||
| EI-267 |
AG-825 |
HER1-2 |
0.35 |
autophos. |
237 |
| EGFR |
19 |
autophos. |
237 |
||
| PDGFR |
40 |
autophos. |
237 |
||
| p210Bcr-Abl |
75 |
unspecified |
238 |
||
| IR |
>100 |
unspecified |
238 |
||
| EI-258 |
AG-879 |
Trk (NGFR) |
10 |
autophos. |
238 |
| EGFR |
>500 |
autophos. |
238 |
||
| PDGFR |
>100 |
autophos. |
238 |
||
| EI-277 |
AG-1478 |
EGFR |
0.003 |
unspecified |
238 |
| HER2-Neu |
>100 |
unspecified |
238 |
||
| PDGFR |
>100 |
unspecified |
238 |
||
| p210Bcr-Abl |
>50 |
unspecified |
238 |
||
| EI-146 |
Erbstatin Analog |
EGFR |
0.8 |
src peptide |
244 |
| EI-147 |
Genistein |
EGFR |
2.6 22 1 |
Autophos. Histone 2B Ang. II |
292 299 |
| v-src |
26 |
Casein |
292 |
||
| c-src |
>50 |
poly-GT |
299 |
||
| v-abl |
39 |
v5-Ang.II |
299 |
||
| PKA |
>100 |
Histone 2A |
292 |
||
| PKC |
15 >100 |
Histone H1 |
299 292 |
||
| EI-232 |
HDBA |
EGFR |
0.011 |
src peptide |
136 |
| c-src |
0.5 |
poly-GT |
135 |
||
| CaMK |
0.2 |
135 |
|||
| EI-247 |
HNMPA |
IR |
100 |
||
| EI-185 |
Lavendustin A |
EGFR |
0.011 |
src peptide |
245 |
| PKA |
>100 |
unspecified |
245 |
||
| PKC |
>100 |
unspecified |
245 |
||
| EI-190 |
Tyrphostin 1 |
EGFR |
>1250 |
poly-GAT |
257 |
| EI-215 |
Tyrphostin 9 |
EGFR |
460 |
poly-GAT |
286 |
| (RG-50872) |
PDGFR |
0.5 |
unspecified |
286 |
|
| EI-191 |
Tyrphostin 23 |
EGFR |
35 |
poly-GAT |
257 |
| (RG-50810) |
42 |
Ang. II |
299 |
||
| PDGFR |
20-30 |
unspecified |
286 |
||
| IR |
1200(Ki) |
poly-GT |
253 |
||
| PKA |
>1000 |
unspecified |
253 |
||
| PKC |
>1000 |
unspecified |
253 |
||
| EI-187 |
Tyrphostin 25 |
EGFR |
3 15 |
poly-GAT Autophos. |
257 253 |
| EI-257 |
Tyrphostin 46 |
EGFR |
9.2 |
unspecified |
286 |
| p56lck |
22 |
enolase |
254 |
||
| PDGF |
50 |
unspecified |
255 |
||
| EI-188 |
Tyrphostin 47 |
EGFR |
2.4 |
poly-GAT |
257 |
| (RG-50864) |
IR |
640(Ki) |
poly-GT |
253 |
|
| PDGFR |
3.5 |
unspecified |
286 |
||
| Trk |
>100 |
unspecified |
238 |
||
| p210Bcr-Abl |
6 |
unspecified |
238 |
||
| EI-189 |
Tyrphostin 51 |
EGFR |
0.8 |
poly-GAT |
257 |
Abbreviations used are: EGFR, epidermal growth factor receptor; IR, insulin receptor; PDGFR, platelet-derived growth factor receptor; PKA, cAMP-dependent protein kinase; PKC, protein kinase C; Autophos., autophosphorylation; Ang.II, angiotensin II; V5-Ang.II [Val5]angiotensin II; poly-GAT, Glu-Ala-Tyr (6:3:1) polymer; poly-GT, Glu-Tyr (4:1) polymer; src peptide, src autophosphorylation site peptide.
ANTI-PHOSPHOTYROSINE ANTIBODIES
SA-240
PY20, Anti-Phosphotyrosine, mouse monoclonal antibody
Mouse monoclonal antibody. Clone PY20. Immunogen: Phosphotyrosine conjugated to KLH300,301. Supplied as purified IgG2b, 0.1 mg/ml. Recognizes
phosphotyrosine in proteins (species independent) and as the free amino acid. No
cross-reactivity with phosphoserine or phosphothreonine. This classic anti-phosphotyrosine
antibody is now available in convenient package sizes at low cost. High concentrations of
salt (>0.2 M) and divalent cations (>1 mM) may inhibit antibody interaction.
Applications: WB 1 µg/ml, IC, IH and IP.
25 µg
5 x 25 µg
SA-241
PY20, Agarose Beads
Anti-phosphotyrosine, clone PY20, covalently coupled to 4% cross-linked agarose beads.
This product is optimized for binding of phosphotyrosine containing proteins with minimal
non-specific binding. Supplied as a 50% slurry (1 ml total volume) at a conjugation ratio
of 8-10 mg of PY20 per ml of packed beads. Application: Immunoprecipitation. Storage: 4°C
0.5 ml
5 x 0.5 ml
SA-242
PY20, Fluorescein Conjugate
Anti-phosphotyrosine, clone PY20, conjugated to fluorescein. Supplied at 0.1 mg/ml IgG2b,
at a fluorescein/antibody molar ratio of ~4. Application: Immunofluorescence. Storage:
4°C
100 µl
SA-243
PY20, Biotin Conjugate
Anti-phosphotyrosine, clone PY20, conjugated to biotin. Supplied at 0.1 mg/ml IgG2b.
Applications: IH, IC. Storage: 4°C
100 µg
SA-154
Anti-Phosphotyrosine
Rabbit polyclonal antibody. Immunogen: Phosphotyrosine conjugated to KLH. Supplied as IgG,
affinity purified against immobilized phosphotyrosine. Applications: WB 1-5 µg/ml, ELISA.
100 µg
REFERENCES
1. Y. Nishizuka Nature 1988 334 661
2. R.M. Bell and D.J. Burns J. Biol. Chem. 1991 266 4661
3. M. Liyanage et al. Biochem. J. 1992 283 781
4. K. Ogita et al. Proc. Natl. Acad. Sci. USA 1992 89 1592
5. H. Leibersperger et al. J. Biol. Chem. 1990 265 16108
6. H. Koide et al. Proc. Nat. Acad. Sci. USA 1992 89 1149
7. T.C. Saido et al. Biochemistry 1990 31 482
8. D. Schaap and P. Parker J. Biol. Chem. 1990 265 7301
9. A. Toker et al. J. Biol. Chem. 1994 269 32358
10. N. Bacher et al. Mol. Cell. Biol. 1991 11 126
11. S. Osada et al. J. Biol. Chem. 1990 265 22434
12. Y. Nishizuka Science 1992 258 607
13. D.K. Ways et al. J. Biol. Chem. 1992 267 4799
14. H. Nakanishi et al. J. Biol. Chem. 1993 268 13
15. R. Gopalakrishna et al. FEBS Lett. 1992 314 149
16. U. Kikkawa et al. Biochem. Biophys. Res. Commun. 1986 135 636
17. J.R. Woodgett and T. Hunter J. Biol. Chem. 1987 262 4936
18. M.D. Browning et al. Proc. Natl. Acad. Sci. USA 1990 87 1315
19. K.P. Huang and F.L. Huang Biochem. Biophys. Res. Commun. 1986 139 320
20. B. VanRenterghem et al. J. Biol. Chem. 1994 269 24666
21. L-Y. Wang et al. J. Physiol. (Lond.) 1994 475 431
22. H. Hug and T.F. Sarre Biochem. J. 1993 291 329
23. R.M. Epand Analytical Biochemistry 1994 218 241
24. M.G. Kazanietz et al. Mol. Pharmacol. 1993 44 298
25. H. Koide et al. Proc. Natl. Acad. Sci. USA 1992 89 1149
26. P.J. Blackshear. J. Biol. Chem. 1993 268 1501
27. J.M. Graff et al. J. Biol. Chem. 1991 266 14390
28. B. Amess et al. FEBS Lett. 1992 297 285
29. D. Schaap et al. FEBS Lett. 1989 243 351
30. I. Yasuda et al. Biochem. Biophys. Res. Commun. 1990 166 1220
31. Y.J.K. Farrar et al. Biochem. Biophys. Res. Commun. 1991 180 694
32. T. Hunter et al. Nature 1984 311 480
33. C. House and B.E. Kemp Science 1987 238 1726
34. S. Majumdar et al. Biochim. Biophys. Acta 1993 1176 276
35. M. Castagna et al. J. Biol. Chem. 1982 257 7847
36. Y. Nishizuka Nature 1984 308 693
37. Y. Nishizuka Science 1984 225 1365
38. J.E. Niedel et al. Proc. Natl. Acad. Sci. USA 1983 80 36
39. Y. Nishizuka Science 1986 233 305
40. P.M. Blumberg Crit. Rev. Toxicol. 1980 8 153
41. K.R. Gustafson et al. J. Med. Chem. 1992 35 1978
42. W.J. Ryves et al. FEBS Lett. 1991 288 5
43. F.J. Evans et al. Biochem. Soc. Trans. 1991 19 397
44. T.A. Deisher et al. Biochem. Biophys. Res. Commun. 1993 193 1283
45. T. Mori et al. J. Biochem (Tokyo) 1982 91 427
46. K. Kaibuchi et al. J. Biol. Chem. 1983 258 6701
47. W.R. Bishop et al. J. Biol. Chem. 1986 261 6993
48. M. Chuang et al. Am. J. Physiol. 1993 265 C927
49. D.E. Kerr et al. Biochem. Biophys. Res. Commun. 1987 148 776
50. M. Go et al. Biochem. Biophys. Res. Commun. 1987 144 598
51. L.T. Boni et al. J. Biol. Chem. 1985 260 10819
52. T. Shinomura et al. Proc. Natl. Acad. Sci. USA 1991 88 5149
53. J. Meldolesi and M. Magni Trends Pharmacol. Sci. 1991 12 362
54. J.P. Walsh and R.M. Bell Methods. Enzymol. 1992 209 153
55. J.E. Preiss et al. J. Biol. Chem. 1986 261 8597
56. J.E. Preiss et al. Methods. Enzymol. 1987 141 294
57. E.G. Lapetina et al. J. Biol. Chem. 1985 260 1358
58. M. Issandou et al. Biochem. Biophys. Res. Commun. 1988 151 458
59. D. de Chaffoy de Courcelles et al. J. Biol. Chem. 1985 260 15762
60. T. Ohtsuka et al. J. Biol. Chem. 1990 265 15418
61. F. Sakane et al. FEBS Lett. 1989 255 409
62. D. de Chaffoy de Courcelles et al. J. Biol. Chem. 1989 264 3274
63. H. Hennings et al. Carcinogenesis 1987 8 1343
64. Z. Szallasi et al. J. Biol. Chem. 1994 269 2118
65. B.A. Gilbert et al. Biochemistry 1995 34 3916
66. J.D. Winkler et al. J. Org. Chem. 1995 60 1381
67. H. Fujiki and T. Sugimura Adv. Cancer Res. 1987 49 223
68. M. Collins and E. Rozengurt Biochem. Biophys. Res. Commun.1982 104 1159
69. M. Ito et al. Biochemistry 1986 25 4179
70. G Martiny-Baron et al. J. Biol. Chem. 1993 268 9194
71. M. Gschwendt et al. Biochem. Biophys. Res. Commun. 1994 199 93
72. Y. Kashiwada et al. J. Med. Chem. 1994 37 195
73. E. Kobayashi et al. J. Antibiot. 1989 42 1470
74. T. Iida et al. J. Antibiot. 1989 42 1475
75. E. Kobayashi et al. Biochem. Biophys. Res. Commun. 1989 159 548
76. R.F. Bruns et al. Biochem. Biophys. Res. Commun. 1991 176 288
77. J.M. Herbert et al. Biochem. Biophys. Res. Commun. 1990 172 993
78. D. Toullec et al. J. Biol. Chem. 1991 266 15771
79. R. Goldman et al. FEBS Lett. 1992 309 190
80. L.W. Daniel et al. Biochem. Biophys. Res. Commun. 1988 151 291
81. W.J. Van Blitterswijk et al. Lipids 1987 22 842
82. I.M. Kramer et al. J. Biol. Chem. 1989 264 5876
83. D. Meruelo et al. Proc. Natl. Acad. Sci. USA 1988 85 5230
84. K. Tanaka et al. Jpn. J. Cancer Res. (Gann) 1986 77 1107
85. T. Eichholtz et al. J. Biol. Chem. 1993 268 1982
86. K.P. Gupta et al. J. Biol. Chem. 1996 271 2102
87. C.A. O'Brian et al. Invest. New Drugs 1991 9 169
88. N.E. Ward and C.A. O'Brian Biochemistry 1993 32 11903
89. G. Hardie Nature 1988 335 592
90. T.J. Ferro et al. Am J Physiol. 1993 264 L7
91. L.W. Slice and S.S. Taylor J. Biol. Chem. 1989 264 20940
92. A. Enz et al. Anal. Biochem. 1994 216 147
93. H.C. Hemmings et al. J. Biol. Chem. 1990 265 20367
94. D.A. Fruman et al. Proc. Natl. Acad. Sci. USA 1992 89 3686
95. A.M. Pomerantz et al. Proc. Nat. Acad. Sci. USA 1977 74 4261
96. B.E. Kemp et al. Fed. Proc. 1976 35 1384
97. R. Roskowki Methods Enzymol. 1983 99 2
98. J. Aronowski et al. J. Neurochem. 1992 58 1743
99. D.A. Malencik and S.R. Anderson Anal. Biochem. 1983 132 34
100. K.J. Murray et al. Biochem. J. 1990 267 703
101. T. Chijiwa et al. J. Biol. Chem. 1990 265 5267
102. M. Muroi and T. Suzuki Cell Signal. 1993 5 289
103. M. Fujihara et al. J. Biol. Chem. 1993 268 14898
104. K. Yamada and A. Ishii Jpn. J. Pharmacol. 1989 49 263P
105. M. Hagiwara et al. Mol. Pharmacol. 1987 31 523
106. R. Spangler et al. Proc. Natl. Acad. Sci. USA 1989 86 7017
107. T. Asano et al. Br. J. Pharmacol. 1989 98 1091
108. Y. Sasaki and Y. Sasaki Biochem. Biophys. Res. Commun. 1990 171 1182
109. T. Asano et al. J. Pharmacol. Exp. Ther. 1987 241 1033
110. J. Thomas et al. J. Biol. Chem. 1991 266 10906
111. W. Wen and S.S. Taylor J. Biol. Chem. 1994 269 8423
112. H.C. Cheng et al. J. Biol. Chem. 1986 261 989
113. D.R. Knighton et al. Science 1991 253 414
114. B.E. Kemp et al. Methods Enzymol. 1988 159 173
115. D.B. Glass et al. J. Biol. Chem. 1989 264 8802
116. L.H. Botelho et al. Methods Enzymol. 1988 159 159
117. P. Schaap et al. J. Biol. Chem. 1993 268 6323
118. I.D. Grozdova et al. Biochem. Intl. 1992 27 811
119. D.E. Cummings et al. Nature 1996 382 622
120. B.S. Skalhegg et al. J. Biol. Chem. 1992 267 5374
121. A. Keilbach et al. Eur. J. Biochem. 1992 208 467
122. J.D. Corbin and S.O. Doskeland J. Biol. Chem. 1983 258 11391
123. J.L. Colbran et al. J. Biol. Chem. 1992 267 9589
124. D.B. Glass Biochem. J.1983 213 159
125. T.K. Machu et al. Alcohol. Clin. Exp. Res. 1991 15 1040
126. T.K. Machu et al. J. Neurochem. 1993 61 375
127. T.E. Morgan et al. J. Cell Biol. 1993 122 623
128. P.I. Hanson et al. Neuron 1989 3 59
129. Y. Hashimoto and T.R. Soderling Arch. Biochem. Biophys. 1987 252 418
130. H. Mochizuki et al. J. Biol. Chem. 1993 268 9143
131. U.R. Rapp Oncogene 1991 6 495
132. P. Li et al. Cell 1991 64 479
133. T. Force et al. Proc. Nat. Acad. Sci. USA 1994 91 1270
134. O. Miyano et al. J. Biol. Chem. 1992 267 1198
135. T.J. O'Dell et al. Nature 1991 353 558
136. I. Takahashi et al. Biochem. Biophys. Res. Commun. 1989 165 1207
137. H. Tokumitsu et al. J. Biol. Chem. 1990 265 4315
138. H. Minami et al. Biochem. Biophys. Res. Commun. 1994 199 241
139. J. Wei et al. J. Biol Chem. 1996 271 24231
140. G.A. Waymen et al. J. Biol. Chem. 1995 270 5830
141. N. Mamiya et al. Biochem. Biophys. Res. Commun. 1993 195 608
142. R.M. Tombes et al. Cell Growth Differ. 1995 6 1063
143. G. Rassmussen and C. Rassmussen Biochem. Cell Biol. 1995 73 201
144. S. Nakanishi et al. Mol. Pharmacol. 1990 37 482
145. R.J. Colbran et al. J. Biol. Chem. 1988 263 18145
146. R.J. Colbran et al. J. Biol. Chem. 1989 264 4800
147. M.N. Waxham et al. Biochemistry 1993 32 2923
148. M.N. Waxham et al. Brain Res. 1993 609 1
149. M.K. Smith et al. J. Biol. Chem. 1992 267 1761
150. A. Ishida and H. Fujisawa J. Biol. Chem. 1995 270 2163
151. A. Ishida et al. Biochem. Biophys. Res. Commun. 1995 212 806
152. N.E. Erondu and M.B. Kennedy J. Neurosci. 1985 5 3270
153. H.C. McKee et al. J. Neuropathol. Exp. Neurol. 1990 49 49
154. B.B. Molloy and M.B. Kennedy Proc. Natl. Acad. Sci. USA 1991 88 4756
155. P.J. Kennelly et al. J. Biol. Chem. 1987 262 11958
156. B. Kemp and R. Pearson J. Biol. Chem. 1985 260 3355
157. C.H. Michnoff et al. J. Biol. Chem. 1986 261 8320
158. M. Saitoh et al. J. Biol. Chem. 1987 262 7796
159. M. Saitoh et al. Biochem. Biophys. Res. Commun. 1986 140 280
160. G. Inoue et al. J. Biol. Chem. 1993 268 5272
161. N. Begum et al. J. Biol. Chem. 1995 270 709
162. B.E. Kemp et al. J. Biol. Chem. 1987 262 2542
163. D.R. Marshak and D. Carroll Methods Enzymol. 1991 200 134
164. J.K. Klarlund and M.P. Czech J. Biol. Chem. 1988 263 15872
165. E.A. Kuenzel and E.G. Krebs Proc. Natl. Acad. Sci. USA 1985 82 737
166. R. Zandomeni et al. J. Biol. Chem. 1986 261 3414
167. T. Wada et al. Nucleic Acids Res. 1996 24 876
168. I. Schmidt-Spaniol et al. Hybridoma 1992 11 53
169. N. Grankowski et al. Eur. J. Biochem. 1991 198 25
170. M.H. Cobb et al. Cell Regulation 1991 2 965
171. D.J. Robbins et al. J. Biol. Chem. 1993 268 5097
172. C.-F. Zheng and K.L. Guan EMBO J. 1994 13 1123
173. T.A. Bird et al. J. Biol. Chem. 1991 266 22661
174. R. Williams et al. J. Biol. Chem. 1993 268 18213
175. T. Force et al. Proc. Natl. Acad. Sci. USA 1994 91 11270
176. J. Sanghera et al. FEBS Lett. 1990 273 223
177. M. Daeipour et al. J. Immunol. 1993 150 4743
178. I. Clark-Lewis et al. J. Biol. Chem. 1991 266 15180
179. M.H. Cobb and E.J. Goldsmith J. Biol. Chem. 1995 270 14843
180. J.E. Casnellie Methods Enzymol. 1991 200 115
181. E.M. Sale et al. EMBO J. 1995 14 674
182. C.J.M. Robinson et al. Biochem. J. 1996 320 123
183. D.T. Dudley et al. Proc. Natl. Acad. Sci. USA 1995 92 7686
184. D.R. Alessi et al. J. Biol. Chem. 1995 270 27489
185. L. Pang et al. J. Biol. Chem. 1995 270 13585
186. J. M. Kyriakis et al. Nature 1994 369 156
187. B. Derijard et al. Cell 1994 76 1025
188. J. Shim et al. Nature 1996 381 27
189. M. Inagaki et al. Mol. Pharmacol. 1986 29 577
190. H. Hidaka et al. Biochemistry 1984 23 5036
191. M. Inagaki et al. J. Biol. Chem. 1985 260 2922
192. M. Wolf et al. J. Biol. Chem. 1985 260 15718
193. H. Kase et al. Biochem. Biophys. Res. Commun. 1987 142 436
194. K. Yamada et al. Biochem. Biophys. Res. Commun. 1987 144 35
195. Y. Hashimoto et al. Biochem. Biophys. Res. Commun. 1991 181 423
196. L.H. Elliott et al. Biochem. Biophys. Res. Commun. 1990 171 148
197. M.M. Berg et al. J. Biol. Chem. 1992 267 13
198. S.H. Nye et al. Mol. Biol. Cell 1992 3 677
199. P. Tapley et al. Oncogene 1992 7 371
200. E.D. Lloyd and M.W. Wooten J. Neurochem. 1992 59 1099
201. T. Tamaoki et al. Biochem. Biophys. Res. Commun. 1986 135 397
202. H. Matsumoto et al. Biochem. Biophys. Res. Commun. 1989 158 105
203. N. Yanagihara et al. J. Neurochem. 1991 56 294
204. R.J. Fallon Biochem. Biophys. Res. Commun. 1990 170 1191
205. J.A. Badwey et al. Biochem. Biophys. Res. Commun. 1991 178 423
206. J.M. Herbert et al. Biochem. Biophys. Res. Commun. 1990 171 189
207. M. Wolf and M. Baggiolini Biochem. Biophys. Res. Commun. 1988 154 1273
208. I. Kiyoto et al. Biochem. Biophys. Res. Commun. 1987 148 740
209. L. Shi et al. Science 1994 263 1143
210. S.L. Pelech et al. Proc. Natl. Acad. Sci. USA 1986 83 5968
211. C. House et al. J. Biol. Chem. 1987 262 772
212. K. Tobe et al. J. Biol. Chem. 1992 267 21089
213. D.L. Brautigan et al. Science 1992 257 1261
214. R. Zinck et al. Mol. Cell. Biol. 1995 15 4930
215. E. Cano et al. J. Cell Sci. 1995 108 3599
216. E. Kardalinou et al. Mol. Cell. Biol. 1994 14 1066
217. S. Jaken et al. Cancer Res. 1983 43 11
218. T.J. Slaga et al. Proc. Natl. Acad. Sci. USA 1980 77 3659
219. T. Hunter and J.A. Cooper Annu. Rev. Biochem. 1985 54 897
220. J.L. Boyer et al. Trends Pharmacol. Sci. 1989 10 360
221. S.G. Rhee et al. Science 1989 244 546
222. G. Carpenter Annu. Rev. Biochem. 1987 56 881
223. Y. Yarden and A. Ullrich Annu. Rev. Biochem. 1988 57 443
224. B. Ek et al. Nature 1982 295 419
225. U.-H. Kim et al. J. Biol. Chem. 1991 266 1359
226. O.M. Rosen Science 1987 237 1452
227. A. Dhar et al. Mol. Pharmacol. 1990 37 519
228. D.C. Gaudette and B.J. Holub Biochem. Biophys. Res. Commun. 1990 170 238
229. A. Golden and J.S. Brugge Proc. Natl. Acad. Sci. USA 1989 86 901
230. S. Nakamura and H. Yamamura J. Biol. Chem. 1989 264 7089
231. J. Gomez-Cambronero et al. Biochem. Biophys. Res. Commun. 1989 162 1478
232. T. Mustelin et al. Science 1990 247 1584
233. J.M. Trevillyan et al. J. Immunol. 1990 145 3223
234. P.J.L. Lane et al. J. Immunol. 1991 146 715
235. J.J. O'Shea et al. Proc. Natl. Acad. Sci. USA 1991 88 350
236. A. Levitzki et al. Methods Enzymol. 1991 201 370
237. N. Osherov et al. J. Biol. Chem. 1993 268 11134
238. A. Levitzki et.al. Science 1995 267 1782
239. H. Daub et al. Nature 1996 379 559
240. N. Osherov and A. Levitski Eur. J. Biochem. 1994 225 1047
241. Z. Fan et al. J. Cell Biol. 1995 131 235
242. Y. Miyazaki et al. Biochem. Biophys. Res. Commun. 1996 226 542
243. M. Imoto et al. Biochem. Int. 1987 15 989
244. K. Umezawa et al. FEBS Lett. 1990 260 198
245. T. Onoda et al. J. Nat. Prod. 1989 52 1252
246. C.-Y.J. Hsu et al. J. Biol. Chem. 1991 266 21105
247. J.L. Kornyei et al. J. Endocrinol. 1995 146 261
248. D.E. Hu and T.P. Fan Br. J. Pharmacol. 1995 114 262
249. T. Yoneda et al. Cancer Res. 1991 51 4430
250. P.A. Oude Weernink et al. Neurosurgery 1996 38 108
251. N. Sion-Vardy et al. J. Surg. Res. 1995 59 675
252. R.M. Lyall et al. J. Biol. Chem. 1989 264 14503
253. P. Yaish et al. Science 1988 242 933
254. T.R. Burke, Jr. et al. J. Med. Chem. 1993 36 425
255. T.R. Burke, Jr. et al. 204th National ACS Meeting 1992, Conference Proceedings
256. A. Levitzki Biochem. Pharmacol. 1990 40 913
257. A. Gazit et al. J. Med. Chem. 1989 32 2344
258. M.D. Waterfield et al. J. Cell Biol. 1982 20 149
259. W.J. Gullick et al. Cancer Res. 1986 22 285
260. M.F. White and C.R. Kahn J. Biol. Chem. 1994 269 1
261. H. Cho et al. Prot. Science 1993 2 977
262. N.E. Keane et al. Eur. J. Biochem. 1994 226 525
263. S.E. Shoelson et al. Proc. Natl. Acad. Sci. USA 1992 89 2027
264. P. Garcia et al. J. Biol. Chem. 1993 268 25146
265. T. Rordorf-Nikolic et al. J. Biol. Chem. 1995 270 3662
266. R. Saperstein et al. Biochemistry 1989 28 5694
267. K. Baltensperger et al. Proc. Natl. Acad. Sci. USA 1992 89 7885
268. C.R. Faltynek et al. Biochemistry 1995 34 12404
269. Y. Uehara and H. Fukazawa Methods Enzymol. 1991 201 370
270. H. Fukazawa et al. Biochem. Pharmacol. 1991 42 1661
271. T. Satoh et al. J. Biol. Chem. 1992 267 2537
272. R. Weiss and R. Nuccitelli J. Biol. Chem. 1992 267 5608
273. J.M. Oliver et al. J. Biol. Chem. 1994 269 29697
274. J. H. Hanke et al. J. Biol. Chem. 1996 271 695
275. L.J. Pike et al. Proc. Natl. Acad. Sci. USA 1982 79 1443
276. J.E. Casnellie et al. Proc. Natl. Acad. Sci. USA 1982 79 282
277. K. Sato et al. Biochem. Biophys. Res. Commun. 1990 171 1152
278. R.B. Birge and H. Hanafusa Science 1993 262 1522
279. Y. Fukami et al. J. Biol. Chem. 1993 268 1132
280. N. Meydan et al. Nature 1996 379 645
281. H. Dadi et al. Blood 1994 84 1579
282. N. Sharfe et al. Blood 1995 86 2077
283. A. Novogrodsky et al. Science 1994 264 1319
284. G. Sarmay et al. Proc. Natl. Acad. Sci. USA 1994 91 4140
285. C.M. Roifman and G. Wang J. Immunol. 1992 149 1179
286. A. Levitzki and C. Gilon Trends Pharmacol. Sci. 1991 12 171
287. M.C. Bryckaert Exp. Cell Res. 1992 199 255
288. M. Ohmichi et al. Biochemistry 1993 32 4650
289. C. Linassier et al. Biochem. Pharmacol. 1990 39 187
290. J. Markovits et al. Cancer Res. 1989 49 5711
291. N.M. Dean et al. Biochem. Biophys. Res. Commun. 1989 165 795
292. T. Akiyama et al. J. Biol. Chem. 1987 262 5592
293. R.J. Miksicek J. Steroid Biochem. Mol. Biol. 1994 49 153
294. Y. Jing and S. Waxman Anticancer Res. 1995 15 1147
295. M. Kusaka and N. Sperelakis Biochim. Biophys. Acta 1996 1278 1
296. G.E. Bilder et al. Am. J. Physiol. 1991 260 C721
297. H. Terada Biochim. Biophys. Acta 1981 639 225
298. A. Gazit et al. J. Med. Chem. 1991 34 1896
299. J.F. Geissler et al. J. Biol. Chem. 1990 265 22255
300. J.R. Glenny, Jr. et al. J. Immunol. Methods 1988 109 277
301. S. Ruff-Jamison et al. J. Biol. Chem. 1991 266 6607
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