Principle of Assay
Rat Complement C4a ELISA Kit (A79913) employs the sandwich enzyme immunoassay technique for the quantitative measurement of rat Complement C4a in serum, plasma, tissue homogenates, and other biological fluids. An antibody specific for Complement C4a has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Complement C4a present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-Complement C4a Antibody, which binds the captured Complement C4a present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of Complement C4a captured in each well. The concentration of Complement C4a can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
