Human Granzyme B ELISA Kit (A312589) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human Granzyme B in serum, plasma or other biological fluids. An antibody specific for Granzyme B has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the Granzyme B present in each sample is bound to the wells by the immobilized antibody. Biotinylated Anti-Granzyme B Antibody, which also binds the Granzyme B present in each sample, and Streptavidin-HRP, which binds the Biotinylated Anti-Granzyme B Antibody, are added and the microtiter plate is incubated. Following incubation, unbound Biotinylated Anti-Granzyme B Antibody and unbound Streptavidin-HRP are removed by washing, and two substrate solutions are added to the wells. Color develops in proportion to the amount of Granzyme B captured in each well. The color development is stopped by addition of stop solution which changes the color from blue to yellow and the intensity of the color is then measured. The concentration of Granzyme B in the samples can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.