This Progesterone (Pg) BioAssay™ ELISA Kit is a competitive ELISA forthe in vitro quantitative determination of Pg concentrations in serum, plasma and other biological fluids.
Specificity:Recognizes Progesterone (Pg). No significant cross-reactivity or interference with pregnenolone, androstenedione, estradiol.
Test Principle:The microtiter plate provided in this kit has been pre-coated with Goat Anti-Rabbit IgG, make solid-phase secondary antibody. Then add samples, horseradish peroxidase-labeled Progesterone and anti-Progesterone antibody, so as to form a coated secondary antibody anti-Progesterone antibody HRP-labeled Progesterone complex. The amount of bound labeled Progesterone is inversely proportional to that of Progesterone in the samples. The TMB substrate solution is added to each well. The enzyme-substrate reaction is terminated by the addition of stop solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 2nm. The concentration of Progesterone in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Storage and Stability:Unused kit is stable for 6 months after receipt. Store Microtiter Strips, Standard, and Antibody-Biotin at 4°C if kit will be used within 30 days; otherwise store at -20°C until expiration date. Store the rest of the components at 4°C. Protect HRP Conjugate and Substrate Reagent from light. For maximum recovery, centrifuge the original vials after thawing and prior to removing the cap.
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 100ul standard or sample to each well. Incubate 90 minutes at 37°C
3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 90ul Substrate. Incubate 15 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
