The 17-Hydroxyprogesterone (17-OHP) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of 17-OHP in serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.
Test Principle: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.
Storage and Stability: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
Assay Procedure Summary: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.
