Principle of Assay
Human Tryptase ELISA Kit (A78914) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Tryptase in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Tryptase. Standards and samples are added to the wells, where Tryptase binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and an HRP-conjugated detection antibody specific for Tryptase is added to the wells. The detection antibody binds the captured Tryptase 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 Tryptase captured in each well. The concentration of Tryptase is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
