The Vasoactive Intestinal Peptide (VIP) BioAssay(TM) ELISA Kit is a competitive ELISA for the in vitro quantitative determination of VIP concentrations in mouse serum, plasma and other biological fluids. Detection Range: 12.5-800pg/ml Sensitivity: 7.5pg/ml Specificity: Recognizes mouse VIP. No significant cross-reactivity or interference between mouse VIP and analogues was observed. Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <10% Test Principle: This ELISA kit uses the competitive principle. The micro ELISA plate provided in this kit has been pre-coated with VIP. During the reaction, VIP in the sample or standard competes with a fixed amount of VIP on the solid phase supporter for sites on the Biotinylated Detection Ab specific to VIP. Excess conjugate and unbound sample or standard are washed from the plate, and Avidin conjugated to Horseradish Peroxidase (HRP) are added to each microplate well and incubated. Then a 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 VIP in the samples is then determined by comparing the O.D. of the samples to the standard curve. Kit Components: *384831A: Microtiter Strips, 1x96 wells, Pre-coated, ready to use *384831B: Standard, 2x1vial 384831C: Standard Diluent, 1x20ml *384831D: Biotin Detection, 100X, 1x120ul *384831E: HRP Conjugate, 100X, 1x120ul 384831F: Biotin Detection Diluent, 1x14ml 384831G: HRP Diluent, 1x14ml 384831H: Substrate, 1x10ml 384831K: Stop Solution, 1x10ml 384831L: Wash Buffer, 25X, 1x30ml Storage and Stability: Store *384831A, *384831B, *384831D and *384831E at -20C. Store all the other components at 4C. 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 50ul standard or sample to each well. 3. Immediately add 50ul prepared Biotin Working Solution. Incubate 45 minutes at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate. Incubate 15 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
