Epinephrine/Adrenaline (EPI) BioAssay™ ELISA Kit is a quantitative competitive assay for the detection of EPI in serum, plasma, tissue homogenates and other biological fluids.Detection Range7.813-500pg/mlSensitivity<4.688pg/mlPrecisionIntra-assay CV: <8%Inter-assay CV: <10%Assay PrincipleThis ELISA kit employs the quantitative competitive enzyme-linked immunoassay technique, utilizing a microtiter plate provided in this kit has been pre-coated with EPI. Standards and samples are added to the appropriate microtiter plate wells followed by a biotin-conjugated antibody specific to EPI. Streptavidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain EPI, Biotin-conjugated antibody and enzyme-conjugated Streptavidin 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 EPI in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components*355458A: Microtiter Strips, 1x48 wells or 1x96 wells*355458B: Standard, 1x1vial or 2x1 vials355458C: Sample/Standard Dilution Buffer, 1x10ml or 1x20ml355458D: Antibody (Biotin) (Concentrated), 1x30ul or 1x60ul355458E: Antibody Dilution Buffer, 1x5ml or 1x10ml355458F: Streptavidin (HRP) (SABC), 1x60ul or 1x120ul355458G: SABC Dilution Buffer, 1x5ml or 1x10ml355458H: TMB Substrate, 1x5ml or 1x10ml355458J: Stop Solution, 1x5ml or 1x10ml355458K: Wash Buffer, 25X, 1x15ml or 1x30mlStorage and StabilityStore unopened *355458A at 4ºC; store at -20ºC once opened. Store unopened *355458B at 4°C; once reconstituted, store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Kit is stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vials after thawing and prior to removing the cap.Assay Summary1. Wash plate 2 times before adding standards and samples to wells.2. Add 50ul standard or sample to each well.3. Immediately add 50ul Antibody-Biotin working solution to each well and incubate for 45 minutes at 37°C4. Aspirate and wash 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate and wash 5 times.7. Add 90ul TMB substrate. Incubate 15-20 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.

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