Principle of Assay
Mouse Grp75/MOT ELISA Kit (A327035) employs the sandwich enzyme immunoassay technique for the quantitative measurement of mouse Grp75/MOT in serum, plasma, and other biological fluids. An antibody specific for Grp75/MOT has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Grp75/MOT 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-Grp75/MOT Antibody, which binds the captured Grp75/MOT present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, 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 Grp75/MOT captured in each well. The concentration of Grp75/MOT can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
