Principle of Assay
Bovine Creatine Kinase MM ELISA Kit (A3955) employs the sandwich enzyme immunoassay technique for the quantitative measurement of bovine Creatine Kinase MM in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. An antibody specific for Creatine Kinase MM has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Creatine Kinase MM 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-Creatine Kinase MM Antibody, which binds the captured Creatine Kinase MM 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 Creatine Kinase MM captured in each well. The concentration of Creatine Kinase MM can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
