The Dopamine (DA) BioAssay™ ELISA Kit is a competitive ELISA for the in vitro quantitative determination of DA concentrations in serum, plasma and other biological fluids.||Detection Range:|31.25-2000pg/ml||Sensitivity:|18.75pg/ml||Precision:|Intra-Assay %CV: <10%|Inter-Assay %CV: <10%||Specificity:|Recognizes DA. No significant cross-reactivity or interference between DA and analogues was observed.||Test Principle:|The micro ELISA plate provided in this kit has been pre-coated with DA. During the reaction, DA in the sample or standard competes with a fixed amount of DA on the solid phase supporter for sites on the Biotinylated Detection Ab specific to DA. 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 DA in the samples is then determined by comparing the O.D. of the samples to the standard curve.||Kit Components:|*529972A: Microtiter Plate, 1x96 wells, Pre-coated; ready to use|*529972B: Standard, 2x1vial |529972C: Standard Diluent, 1x20ml|*529972D: Biotin Detection, 100X, 1x120ul |*529972E: HRP Conjugate, 100X, 1x120ul |529972F: Biotin Detection Diluent, 1x14ml|529972G: HRP Diluent, 1x14ml|529972H: Substrate, 1x10ml|529972K: Stop Solution, 1x10ml|529972L: Wash Buffer, 25X, 1x30ml ||Storage and Stability:|Store *529972A, *529972B, *529972D and *529972E 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 50ul standard or sample to each well.|3. Immediately add 50ul prepared Biotin Working Solution. Incubate 45 minutes 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.