Principle of Assay
Mouse MCP5 ELISA Kit (Small Sample Volume) (A323198) employs the sandwich Enzyme-Linked Immunoassay technique for the quantitative measurement of mouse in serum, plasma, and other biological fluids. This assay is optimized for limited sample availability, requiring only 25 µL of sample per well. An antibody specific for MCP5 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the MCP5 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-MCP5 antibody, which binds the captured MCP5 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 MCP5 captured in each well. The concentration of MCP5 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
