Principle of Assay
Hamster IgM ELISA Kit (A350676) is a 3 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of hamster IgM in serum, plasma, cell culture supernatant, cell or tissue lysate, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for IgM. Standards and samples are added to the wells, where IgM binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for IgM is added to the wells. The detection antibody binds the captured IgM to form an antibody-antigen-antibody sandwich complex. After further incubation and washing, TMB substrate solution is added to visualize the HRP enzymatic reaction. This produces a blue-coloured product that changes to yellow following addition of acidic stop solution. The intensity of the yellow colour is directly proportional to the amount of IgM captured in each well. The concentration of IgM is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
