Description
For samples: Serum, plasma, cell culture supernatants, body fluid and tissue homogenate
INTENDED USE This HSPA1A ELISA kit is a 1.5 hour solid-phase ELISA designed for the quantitative determination of Canine HSPA1A. This ELISA kit for research use only, not for therapeutic or diagnostic applications!
PRINCIPLE OF THE ASSAY HSPA1A ELISA kit applies the competitive enzyme immunoassay technique utilizing a monoclonal anti-HSPA1A antibody and an HSPA1A-HRP conjugate. The assay sample and buffer are incubated together with HSPA1A-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 HSPA1A concentration since HSPA1A from samples and HSPA1A-HRP conjugate compete for the anti-HSPA1A antibody binding site. Since the number of sites is limited, as more sites are occupied by HSPA1A from the sample, fewer sites are left to bind HSPA1A-HRP conjugate. A standard curve is plotted relating the intensity of the color (O.D.) to the concentration of standards. The HSPA1A concentration in each sample is interpolated from this standard curve