The Anti-Smooth Muscle Antibody （ASMA） BioAssay ELISA Kit （Human） is a quantitative sandwich assay for the detection of ASMA in human serum，plasma and other biological fluids.Detection Range：7.813-500ng/mlSensitivity：<4.688ng/mlPrecision： Intra-Assay CV： <8%Inter-Assay CV： <10%Assay Principle：The microtiter plate provided in this kit has been pre-coated with antigen （Myosin）. Standards and samples are added to the appropriate microtiter plate wells followed by biotinylated antigen （biotinylated Myosin）. Streptavidin conjugated to horseradish peroxidase （HRP） is added to each microplate well and incubated. TMB substrate solution is added，after which the enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the resulting color measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of ASMA in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components：*369761A： Microtiter Strips，8x12 wells.*369761B： Standard，2 vials369761C： Sample/Standard Dilution Buffer，1x20ml369761D： Myosin-Biotin （Concentrated），1x120ul369761E： Antigen Dilution Buffer，1x10ml*369761F： Streptavidin-HRP Conjugate （SABC），1x120ul369761G： SABC Dilution Buffer，1x10ml*369761H： TMB Substrate，1x10ml369761J： Stop Solution，1x10ml369761K： Wash Buffer （25X），1x30mlStorage and Stability：Store unopened *369761A at 4ºC; store at -20ºC once opened. Store unopened *369761B at 4°C; once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store *369761F and *369761H in the dark at 4°C. Store all other components at 4°C. Unused 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 Summary：1. Wash plate 2 times before adding standards，samples and control （zero） to wells.2. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C 3. Aspirate the liquid and wash plate 2 times.3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C4. Aspirate the liquid and wash plate 3 times.5. Add 100ul SABC working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate the liquid and wash plate 5 times.7. Add 90ul TMB substrate. Incubate 15-30 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately.9. Calculate results.
