Calcitonin ELISA Cat.-No. : RE 530 01

Size : 12 x 8

Short Instruction Storage : 2 - 8 °C

 

 

1. Preparation for assay

 

Reconstitute standards and control with 0.25 ml (zero standard 0.5 ml) distilled water. Allow the vials to stand for 15 minutes. After reconstitution the standards and control should be stored frozen at - 20 °C.

Dilute the required amount of anti hCT antibodies freshly with assay buffer 1 to 101 (e.g. 40 µl anti hCT antibodies + 4 ml assay buffer).

Dilute the required amount of enzyme conjugate freshly with assay buffer 1 to 101 (e.g. 40 µl enzyme conjugate + 4 ml assay buffer).

Mix the solutions of reagent 1, reagent 2 and reagent 3 in the ratio 1 : 1 : 1 (e.g. 1.5 ml of each) to obtain the substrate solution. Prepare the required amount only directly before use.

Dilute the wash buffer concentrate with distilled water 1 to 10 (e.g. 10 ml concentrate + 90 ml distilled water). Ready to use wash buffer is stable for 6 weeks when stored at 2 - 8 °C.

 

 

2. Specimen Collection and Storage

 

Sample: Serum, EDTA plasma

Storage up to 4 hours: 2 - 8 °C

for longer period: - 20 °C

Samples with values above the highest standard have to be diluted with zero standard.

 

 

3. Assay Procedure

 

Allow any refrigerated or frozen reagents or samples to warm up to room temperature.

Patient results are read from the graph constructed from the standards. Only diluted samples have to be corrected by the dilution factor.

 

 

 

 

 

  • pipet 100 µl assay buffer into each well

  • pipet 20 µl standards, controls or sample into the wells, cover plate with sealing tape
    		•

  • incubate for 3 - 4 hours at room temperature (18 - 24 °C) on a microtiter plate shaker (500 rpm)
    		•

  • *decant supernatant, wash 4x with 250 µl diluted wash buffer remove any residual
    		•

  • pipet 100 µl anti hct biotin into each well, cover plate with sealing tape
    		•

  • incubate overnight (10 - 16 hours) at 2 - 8 °C

  • *decant supernatant, wash 4x with 250 µl diluted wash buffer remove any residual
    		•

  • pipet 100 µl enzyme conjugate into each well, cover plate with sealing tape
    		•

  • incubate for 2 hours at room temperature
    (18 - 24 °C) on a microtiter plate shaker (500 rpm)
    		•

  • *decant supernatant, wash 4x with 250 µl diluted wash buffer remove any residual
    		•

  • **pipet 100 µl substrate solution in each well
    		•

  • incubate for 10 ± 2 minutes at room temperature (18 - 24 °C) on a microtiter plate shaker (500 rpm)
    		•

  • **add 50 µl stop solution and shake plate for homogenising
    		•

  • measure absorbance at 490 nm within 1 hour

    • * Wash procedure is essential for the assay results.

    ** Avoid any air bubbles at the pipetting step. Stop solution should be pipetted in the same time schedule as substrate solution.

     

    4. Expected values

     

    Serum/plasma < 20 pg/ml

     

     

    Caution: For comprehensive information about the test procedure please refer to the detailed instructions for use available on the Internet (http://www.IBL-Hamburg.com) or upon request from IBL in Hamburg.

    08.Aug.98/dhm

    Calcitonin ELISA

    Cat.-No. RE 530 01

     

     

    Assay advantages for the IBL assay

     

     

    Potential customers

    University-labs, Private-labs

     

     

    Clinical Applications

    Diagnosis of medullary thyroid carcinoma (MTC).

    Follow up for recurrence in patients with previous MTC.

    Diagnosis of preclinical MTC patients.

     

    25.09.98/dhm