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蚓
Supplied at 35 痢/ml. Specific activity: >1 痠ol phosphate/min/mg enzyme using
histone H1 as substrate at pH 7.4 and 22蚓. Consists primarily of a,
b and g isotypes15,16,17,18.
1.75 痢
SE-133
Protein kinase C, catalytic fragment (PKM) (from rat brain)
>95% by SDS-PAGE, MW=55 kDa, Storage: -70蚓
Supplied at 10 痢/ml. Specific activity: 0.8 痠ol phosphate/min/mg enzyme using histone
H1 as substrate at pH 7.4 and 30蚓. 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 痢
5 x 0.25 痢
SE-143
Protein kinase C alpha (PKCa) (human, recombinant)
>95% by SDS-PAGE, MW=76.8 kDa, (EC 2.7.1.37) Storage: -70蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.5 痠ol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2,
100 然 CaCl2, 100 痢/ml phosphatidylserine (PS), 20 痢/ml
diacylglycerol (DAG), 0.2 mg/ml histone H1, 100 然 ATP and 0.03% Triton X-100 at pH 7.4
and 30蚓. 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 痢
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蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~0.9 痠ol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2,
100 然 CaCl2, 100 痢/ml phosphatidylserine (PS), 20 痢/ml
diacylglycerol (DAG), 0.2 mg/ml histone H1, 100 然 ATP and 0.03% Triton X-100 at pH 7.4
and 30蚓. 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 痢
SE-146
Protein kinase C gamma (PKCg) (human, recombinant)
>95% by SDS-PAGE, MW=78.4 kDa, (EC 2.7.1.37) Storage: -70蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 痠ol phosphate/min/mg protein
using histone H1 as substrate. Reaction conditions are 10 mM MgCl2, 100 然
CaCl2, 100 痢/ml phosphatidylserine (PS), 20 痢/ml diacylglycerol (DAG), 0.2 mg/ml
histone H1, 100 然 ATP and 0.03% Triton X-100 at pH 7.4 and 30蚓. 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 痢
SE-147
Protein kinase C delta (PKCd) (human, recombinant)
>95% by SDS-PAGE, MW=77.5 kDa, (EC 2.7.1.37) Storage: -70蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.5 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 痠ol 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 痢/ml phosphatidylserine (PS), 20 痢/ml diacylglycerol (DAG),
0.1 mg/ml substrate, 100 然 ATP and 0.03% Triton X-100 at pH 7.4 and 30蚓. May be used
to phosphorylate target substrates of PKCd and to study enzyme
regulation and kinetics24,25.
10 痢
SE-148
Protein kinase C epsilon (PKCe) (human, recombinant)
>95% by SDS-PAGE, MW=83.5 kDa, (EC 2.7.1.37) Storage: -70蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.1 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~1.2 痠ol 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 痢/ml phosphatidylserine (PS), 20 痢/ml
diacylglycerol (DAG), 0.1 mg/ml substrate, 100 然 ATP and 0.03% Triton X-100 at pH 7.4
and 30蚓. May be used to phosphorylate target substrates of PKCe
and to study enzyme regulation and kinetics24,25.
10 痢
SE-149
Protein kinase C zeta (PKCz) (human, recombinant)
>95% by SDS-PAGE, MW=67.7 kDa, (EC 2.7.1.37) Storage: -70蚓
Highly active enzyme expressed in a baculovirus expression system. Supplied at 0.2 mg/ml
in 50% glycerol/stabilizing buffer. Specific activity: ~0.4 痠ol 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 痢/ml phosphatidylserine (PS), 0.1 mg/ml
substrate, and 100 然 ATP at pH 7.5 and 30蚓. May be used to phosphorylate target
substrates of PKCz and to study enzyme regulation and
kinetics23,24
10 痢
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蚓
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蚓
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蚓
Sequence is from myelin basic protein (4-14)30. Specific
substrate for a-,b-, g-isoforms of PKC (Km=7 然) 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蚓
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蚓
PKC pseudosubstrate sequence but Ala to Ser-25 substitution makes it a good substrate (Km=0.2
然) 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蚓
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蚓
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蚓
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蚓
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蚓
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蚓
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蚓
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蚓
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蚓
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蚓54. Specific activity for ceramide phosphorylation is
~0.5 mmol ceramide/min/mg protein at 25蚓. 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蚓
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蚓
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蚓
PKC activator.
20 mg
100 mg
DG-125
1-Oleoyl-2-acetyl-sn-glycerol (OAG)
98%, MW=398.6 [86390-77-4] Storage: -20蚓
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蚓
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蚓
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蚓
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 然) and in
isolated membranes (IC50=6.7 然)59,60. In
porcine thymus cytosol it inhibited the 80 kDa DAG kinase (IC50=10 然) 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蚓
R 59949 is a more potent analog of R 59022. It inhibits human platelet DAG kinase in
intact cells (IC50=0.12 然) and in isolated membranes (IC50=0.3
然)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蚓
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 痢
5 x 10 痢
PE-190
Farnesyl thiotriazole
98%, MW=305.5 [156604-45-4] Storage: -20蚓
A structurally unique activator of PKC in whole cells or in purified enzyme preparations
(KD=2.5 然) 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蚓
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蚓
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蚓
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蚓
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蚓
Calphostin C is a novel, potent and highly selective inhibitor of protein kinase C (IC50=0.05
然)73,74. Inhibition is via interaction with the regulatory
domain75 and is light dependent76.
See Table 4 for selectivity data.
100 痢
1 mg
EI-225
Chelerythrine chloride
98%, MW=383.8 [3895-92-9] Storage: 0蚓
A potent and selective inhibitor of protein kinase C77 (IC50=0.66
然). See Table 4 for selectivity data.
5 mg
25 mg
EI-246
GF 109203X
98%, MW=412.5 [133052-90-1] Storage -20蚓
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
然 (inhibits a, bI, bII and g subtypes with similar potency);
PKA, IC50=2.0 然; phosphorylase kinase, IC50=0.7 然; insulin, EGF
and PDGF receptor tyrosine kinases are not inhibited at concentrations up to 50 然78.
1 mg
5 x 1 mg
EI-149
(?1-O-Hexadecyl-2-O-methylglycerol
99%, MW=330.6, Storage: 0蚓
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蚓
A potent and selective inhibitor of protein kinase C136 (IC50=3.4
然) 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蚓
Inhibits protein kinase C. Completely inhibits transformation of BALB/3T3 cells at 30
痢/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蚓
Pseudosubstrate sequence from PKCa and b.
Highly specific inhibitor which is N-terminal myristoylated to allow membrane
permeability. IC50=8 然 for TPA activation of MARCKS phosphorylation in
fibroblast primary cultures, and 98% inhibition at 100 然85.
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蚓
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 然)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蚓
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蚓
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 然, PKA=423 然, MLCK=24 然33,34.
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 痢
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 痢
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 痢
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 痢
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 痢
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 痢
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 痢
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 痢
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 痢
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 痢 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蚓
Partially purified preparation, supplied lyophilized. Specific activity: 30-65 U/痢
protein. One unit will add 1 pmol phosphate to dephosphorylated casein per minute at pH
6.5 and 30蚓. 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蚓
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蚓. 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 然. |
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蚓
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蚓
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蚓
Substrate for Ser/Thr protein kinases. Km=4.7 然 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蚓
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 然 for PKA; 223 然
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蚓
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蚓
KT5720 is a hexylester derivative of K-252a and is a selective inhibitor of PKA (Ki=0.056
然)193,104. See Table 4 for
selectivity data.
100 痢
5 x 100 痢
EI-158
H-8
N-[2-(Methylamino)ethyl]-5-isoquinolinesulfonamide?HCl
99%, MW=338.3 [84478-11-5] Storage: 0蚓
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蚓
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廈Cl
98%, MW=327.8 [103745-39-7] Storage: 0蚓
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蚓
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蚓
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蚓
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蚓
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
然)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 痢
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 痞 (~1-2.5 痢) 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 痞
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 痞
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 痢
SP-270
Anti-PKA RIIb (SA-270, -271) control peptide
>90%, MW=1980.0 Storage: -20蚓
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 痢
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 痞/lane. Useful as a Western blotting
control for PKA RIIb antibody (SA-270 and -271, above), or for
use with other antibodies.
200 痞
SW-104
Mouse Brain Extract, Immunoblotting Standard
Whole brain protein extract from mouse. Similar to SW-103 above, but from mouse.
200 痞
PROTEIN KINASE G (PKG)
PKG ENZYME
SE-118
Protein kinase G (from bovine lung)
cGMP-dependent protein kinase
90%, Storage: -70蚓
Specific activity: 1-2.5 kU/痢 protein, 10-20 pmol cGMP bound/痢. 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 然), 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蚓
Sequence derived from bovine lung cGMP specific phosphodiesterase (cG-BPDE). Specific
substrate for PKG, Km=68 然; Km for PKA=320 然, 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蚓
KT5823 is a derivative of K-252a and is a selective inhibitor of PKG (Ki=0.234
然)193. See Table 4 for selectivity data.
100 痢
5 x 100 痢
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廈Cl
See "PKA inhibitors".
P-208
PKG inhibitor
Arg-Lys-Arg-Ala-Arg-Lys-Glu
>97%, MW=943.1 [82801-73-8] Storage: -20蚓
Sequence is histone H2B (29-35) non-phosphorylatable analog (Ser to Ala-32) of substrate
peptide of PKG. Ki =86 然 for PKG; Ki =550 然 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蚓
Composed of 60 and 50 kDa subunits in ratios of 1:3 to 1:4. Supplied at 20 痢/ml.
Specific activity: 0.9 痠ol phosphate/min/mg using synapsin I as substrate at 30蚓.
Standard assay conditions are 50 mM HEPES, pH 7.4, 10 mM MgCl2, 100
然 ATP, 5 mM CaCl2, 30 痢/ml calmodulin (catalog #SE-101) and 0.6
ng/痞 CaM kinase II in a total volume of 50 痞125-127.
0.5 痢
5 x 0.5 痢
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蚓
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
然 for CaMKII; >50 然 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蚓
Phosphorylation site 2 of glycogen synthase containing the R-X-X-S/T consensus
phosphorylation site129. Substrate for CaMKII (Km
=12 然) 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蚓
Sequence from Ca2+/calmodulin-dependent protein kinase II-g
(345-358). Substrate is specific for CaMKIV (Km=8 然) 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蚓
Inhibits Ca2+/calmodulin kinase II (IC50=0.2 然)135.
Also inhibits EGF receptor tyrosine kinase (IC50=0.044 然)136
and pp60c-src(+) kinase (IC50=0.5 然)135.
2 mg
10 mg
EI-230
KN-62
98%, MW=721.9 [127191-97-3] Storage: 0蚓
KN-62 is a potent and selective inhibitor of Ca2+/calmodulin kinase II (Ki=0.9
然). 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 然 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蚓
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蚓
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 痢
5 x 100 痢
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蚓
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 然, 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蚓
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
然) 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蚓
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 然 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蚓
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 痢/ml,
IC 10 痢/ml.
100 痢
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蚓
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 然 for rabbit skeletal muscle MLCK, 7.5 然 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蚓
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 然
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廈Cl
99%, MW=452.7 [109376-83-2] Storage: 0蚓
Potent and selective inhibitor of MLC kinase (Ki=0.3 然)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廈Cl
99%, MW=361.3 [105637-50-1] Storage: 0蚓
Selective inhibitor of MLC kinase (Ki=3.8 然)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蚓
Sequence from skeletal muscle myosin light chain kinase (MLCK) (342-352) pseudosubstrate
site. Inhibits MLCK (Ki =1 然 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蚓
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蚓163.
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蚓
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蚓
DRB is a potent and specific inhibitor of casein kinase II (CKII, IC50~6 然).
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
痢/ml.
100 痢
MAP KINASE (MAPK)
MAPK ENZYME
SE-137
MAP Kinase, activated (rat, recombinant)
>95% by SDS-PAGE, MW=42 kDa, Storage: -70蚓
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 痠ol phosphate/min/mg at 30蚓 using myelin basic protein (MBP) as
substrate. Useful to study enzyme regulation, kinetics, and phosphorylate target
substrates170-172.
5 痢
5 x 5 痢
SE-139
MAP Kinase, nonactivated (rat, recombinant)
>95% by SDS-PAGE, MW=42 kDa, Storage: -70蚓
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蚓 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 痢
5 x 20 痢
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蚓
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蚓
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蚓
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蚓.
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 然,
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 然181. It depleted MAP kinase >90%
in rat aortic smooth muscle cells (DOTAP complex, 5-10 然)182.
A 10 然 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蚓.
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蚓
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蚓 using GST-c-jun (1-79) fragment (catalog #SE-151, below)
as substrate. Useful to study enzyme regulation, kinetics, and phosphorylate target
substrates.
20 痢
5 x 20 痢
SE-151
C-JUN (1-79), c-jun kinase substrate (rat, recombinant)
>95% by SDS-PAGE, MW=37 kDa, Storage: -70蚓
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 痢 per assay187,188.
Supplied at 1 mg/ml.
250 痢
5 x 250 痢
SER/THR KINASES - BROAD SPECIFICITY INHIBITORS
EI-145
A-3
N-(2-Aminoethyl)-5-chloronaphthalene-1-sulfonamide廈Cl
98%, MW=321.23, Storage: 0蚓
Inhibits PKA (Ki=4.3 然), PKG (Ki= 3.8 然), MLCK (Ki=7.4
然) and PKC (Ki=47 然)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蚓
Inhibits PKA (Ki=3.0 然), PKG (Ki=5.8 然), MLCK (Ki=97
然) and PKC (Ki=6 然)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蚓
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蚓
Inhibits PKA (Ki=0.018 然), PKG (Ki=0.020 然), MLCK (Ki=0.017
然), PKC (Ki=0.025 然)193,194, CaMK (Ki=0.0018
然)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 痢
1 mg
EI-234
K-252b
99%, MW=453.5 [99570-78-2] Storage: 0蚓
Inhibits PKA (Ki=0.09 然), PKG (Ki=0.1 然), MLCK (Ki=0.147
然) and PKC (Ki=0.02 然)193. See Table 4.
100 痢
1 mg
EI-156
Staurosporin
99%, MW=466.5 [62996-74-1] Storage: 0蚓
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 痢
1 mg
NEW LOWER PRICE
TABLE 4. SPECIFICITIES OF SELECTED SER/THR KINASE INHIBITORS.
[Inhibition as Ki or (IC50), concentration in 然]
| 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蚓
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)微i4 (ATPg-S)
>85%, MW=547.0 [93839-89-5] Storage: -20蚓
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蚓
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蚓
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蚓
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蚓.
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 然) and EGF-dependent
cell growth (IC50=6 然)236. It selectively
inhibits HER1 (EGF receptor) vs. HER1-2 receptor autophosphorylation. HER1: IC50=1.1
然; HER1-2: IC50=45 然237. 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蚓
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 然239. 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 然)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 然)136
and pp60c-src(+) kinase (IC50=0.5 然)135.
Also inhibits Ca2+/calmodulin kinase II (IC50=0.2 然)135.
EI-185
Lavendustin A
98%, MW=381.4 [125697-92-9] Storage: 0蚓
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
然) and PDGF receptor tyrosine kinases and is long acting. It inhibits EGF-stimulated
HER14 cell proliferation (IC50=1 然) 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 然)
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蚓
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 痢
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 痢
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蚓
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蚓
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蚓
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蚓
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蚓
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蚓
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 然)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蚓
HNMPA is a membrane impermeable inhibitor of insulin receptor tyrosine kinase activity (IC50=100
然)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蚓
HNMPA-(AM)3 is a cell permeable analog of HNMPA. It inhibits insulin receptor
tyrosine kinase activity (IC50=100 然) and insulin-stimulated glucose
oxidation in isolated rat adipocytes (IC50=10 然). It has no effect on PKA (at
concs. up to 1 mM) or PKC (at concs. up to 420 然)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蚓
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 痢
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 然274.
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蚓
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蚓
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蚓
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蚓
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
然 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 痞
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 痢/ml, IP (active in
immune-complex kinase assays), IH.
100 痢
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 然. EGFR, Her-2/neu receptor or PDGF-R are not
inhibited at 100 然283. Inhibits tyrosine kinase-dependent
B cell receptor signalling284,285.
5 mg
25 mg
EI-229
AG-370
99%, MW=259.3 Storage: 0蚓
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 然) 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蚓
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 然)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 痢/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
然)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(然) |
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 |
|