Principle of Assay
Canine Anti-KLH IgM ELISA Kit (A319512) employs the Capture ELISA technique for the quantitative measurement of canine Anti-KLH IgM in serum, plasma, tissue homogenates, and other biological fluids. A capture antibody specific for Anti-KLH IgM is pre-coated onto a 96-well microtiter plate. Samples and standards are added to the wells, allowing Anti-KLH IgM to bind to the immobilized antibody. After washing away unbound substances, a detection reagent such as a biotin-labeled or HRP-conjugated antigen, antibody, or streptavidin complex is added to form a measurable complex. Following incubation and washing, TMB substrate is added to visualize the enzymatic reaction, producing a blue-coloured product that changes to yellow after addition of acidic stop solution. The intensity of yellow is proportional to the amount of Anti-KLH IgM captured in each well. The concentration of Anti-KLH IgM can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
