Defensin Beta 2 (DEFb2) BioAssay(TM) ELISA Kit (Human)

The Defensin, beta 2 (DEFb2) BioAssay(TM) ELISA Kit (Human) is a sandwich enzyme immunoassay for the in vitro quantitative measurement of DEFb2 in human serum, plasma, cell lysates, tissue homogenates, cell culture supernatants and other biological fluids. Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to DEFb2. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to DEFb2. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain DEFb2, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of DEFb2 in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *024607A: Microtiter Plate, 1x96 wells. Pre-coated, ready to use., *024607B: Standard, 2x1vial , 024607C: Standard Diluent, 1x20ml, *024607D: Detection Reagent A, 1x120ul , *024607E: Detection Reagent B, 1x120ul , 024607F: Assay Diluent A, 1x12ml, 024607G: Assay Diluent B, 1x12ml, 024607H: TMB Substrate, 1x9ml, 024607K: Stop Solution, 1x6ml, 024607L: Wash Buffer, 30X, 1x20ml , Storage and Stability: Store *024607A, *024607B, *024607D and *024607E 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 date of the kit. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap. Detection Range: 31.2 - 2,000pg/ml, Sensitivity: <13.2pg/ml, Precision: Intra-Assay: CV<10%, Inter-Assay: CV<12%, Assay Procedure Summary: 1. Prepare all reagents, samples and standards., 2. Add 100ul standard or sample to each well. Incubate for 1 hour at 37°C., 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C., 4. Aspirate and wash 3 times., 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C., 6. Aspirate and wash 5 times., 7. Add 90ul TMB Substrate. Incubate 10-20 minutes at 37°C., 8. Add 50ul Stop Solution. Read at 450nm immediately.