Wingless Type MMTV Integration Site Family, Member 3A (WNT3A) BioAssay(TM) ELISA Kit (Mouse)

Wingless Type MMTV Integration Site Family, Member 3A (WNT3A) BioAssay(TM) ELISA Kit (Mouse) is a sandwich enzyme immunoassay for in vitro quantitative measurement of WNT3A in mouse serum, plasma and other biological fluids. Detection Range: 15.6-1000pg/ml, Sensitivity: 5.4pg/ml, Intra-Assay CV: <10%, Inter-Assay CV: <12%, Principle of the Test: The wells of the microtiter strips provided in this kit have been pre-coated with an antibody specific to WNT3A. Standards or samples are then added to the appropriate wells with a biotin-conjugated antibody specific to WNT3A. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. After TMB substrate solution is added, only those wells that contain WNT3A, 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 WNT3A in the samples is then determined by comparing the O.D. of the samples to the standard curve. Kit Components: *153259A: Microtiter Strips, 1x96 wells, Pre-coated, ready to use, *153259B: Standard, 2x1 vial, 153259C: Standard Diluent, 1x20ml, *153259D: Detection Reagent A (green), 1x120ul, *153259E: Detection Reagent B (red), 1x120ul, 153259F: Assay Diluent A, 1x12ml, 153259G: Assay Diluent B, 1x12ml, 153259H: TMB Substrate, 1x9ml, 153259K: Stop Solution, 1x6ml, 153259L: Wash Buffer, 30X, 1x20ml, Precaution: The Stop Solution (153259K) included for use in this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *153259A, *153259B, *153259D and *153259E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months after receipt. 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. Assay Procedure Summary: 1. Prepare all reagents, samples and standards., 2. Add 100ul standard or sample to each well. Incubate 2 hours 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 Substrate Solution. Incubate 15-25 minutes at 37°C., 8. Add 50ul Stop Solution. Read at 450nm immediately.