Principle of Assay
Canine Anti-KLH IgM ELISA Kit (A319512) is a capture Enzyme-Linked Immunosorbent Assay (cELISA) designed for the in vitro quantitative determination of canine Anti-KLH IgM in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with a capture antibody specific for Anti-KLH IgM. Standards and samples are added to the wells, where Anti-KLH IgM present in each sample binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an assay-specific detection reagent is added to the wells. This detection reagent binds the captured Anti-KLH IgM to form a capture immune complex. After further incubation and washing, TMB substrate solution is added to visualize the enzyme-mediated 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 Anti-KLH IgM captured in each well. The concentration of Anti-KLH IgM is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
