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