Description
For samples: Serum, plasma, cell culture supernatants, body fluid and tissue homogenate
INTENDED USE This ?-SMA ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Bovine ?-SMA. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
PRINCIPLE OF THE ASSAY ?-SMA ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-?-SMA antibody and an ?-SMA-HRP conjugate. The assay sample and buffer are incubated together with ?-SMA-HRP conjugate in pre-coated plate for one hour. After the incubation period, the wells are decanted and washed five times. The wells are then incubated with a substrate for HRP enzyme. The product of the enzyme-substrate reaction forms a blue colored complex. Finally, a stop solution is added to stop the reaction, which will then turn the solution yellow. The intensity of color is measured spectrophotometrically at 450nm in a microplate reader. The intensity of the color is inversely proportional to the ?-SMA concentration since ?-SMA from samples and ?-SMA-HRP conjugate compete for the anti-?-SMA antibody binding site. Since the number of sites is limited, as more sites are occupied by ?-SMA from the sample, fewer sites are left to bind ?-SMA-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The ?-SMA concentration in each sample is interpolated from this standard curve