Principle of Assay
Mouse Complement C5a ELISA Kit (A323211) employs the sandwich Enzyme-Linked Immunoassay technique for the quantitative measurement of mouse Complement C5a in serum, plasma, and other biological fluids. An antibody specific for Complement C5a has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Complement C5a 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 C5a Antibody, which binds the captured Complement C5a present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Avidin 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 C5a captured in each well. The concentration of Complement C5a can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
