246.9-20,000pg/ml

96.5pg/ml

Intra-Assay CV: <10% Inter-Assay CV: <12%

This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for 17-OHP has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled 17-OHP and unlabeled 17-OHP (standards or samples) with the pre-coated antibody specific for 17-OHP. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is inversely proportional to the concentration of 17-OHP in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of 17-OHP in the sample.

*023022A: Microtiter Plate, 96 wells, Pre-coated, ready to use *023022B: Standard, 2x1vial 023022C: Standard Diluent, 1x20ml *023022D: Detection Reagent A, 1x120ul *023022E: Detection Reagent B, 1x120ul 023022F: Assay Diluent A, 1x12ml 023022G: Assay Diluent B, 1x12ml 023022H: TMB Substrate, 1x9ml 023022K: Stop Solution, 1x6ml 023022L: Wash Buffer, 30X, 1x20ml

Store *023022A, *023022B, *023022D and *023022E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration d

1. Prepare all reagents, samples and standards. 2. Add 50ul standard or sample to each well, then add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C. 3. Aspirate and wash 3 times. 4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C. 5. Aspirate and wash 5 times. 6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37°C. 7. Add 50ul Stop Solution. Read absorbance at 450nm immediately.