Principle of Assay
Human Fc epsilon RI/FCER1A ELISA Kit (A75401) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Fc epsilon RI/FCER1A in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Fc epsilon RI/FCER1A. Standards and samples are added to the wells, where Fc epsilon RI/FCER1A binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for Fc epsilon RI/FCER1A is added to the wells. The detection antibody binds the captured Fc epsilon RI/FCER1A 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 Fc epsilon RI/FCER1A captured in each well. The concentration of Fc epsilon RI/FCER1A is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
