Description
The Eagle Biosciences Human Anti-Mouse Antibody (HAMA) ELISA Assay Kit is designed, developed and produced for the quantitative measurement of HAMA in serum and plasma samples. The assay utilizes the two-site “sandwich” technique with two selected antibodies that bind to HAMA. Assay standards, controls and patient samples are directly added to wells of a microplate that is coated with murine IgG. After the first incubation period, the HAMA binds to the murine IgG on the wall of microtiter well and unbound proteins in each microtiter well are washed away. Then a horseradish peroxidase (HRP) labeled murine IgG is added to each microtiter well and a “sandwich” of “murine IgG HAMA – murine IgG” is formed. The unbound HRP conjugated murine IgG is removed in the subsequent washing step. For the detection of this immunocomplex, the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the immunocomplex bound to HAMA on the wall of the microtiter well is directly proportional to the amount of HAMA in the sample. A standard curve is generated by plotting the absorbance versus the respective HAMA concentration for each standard on point-to-point, cubical scales or 4 parameter curve fit. The concentration of HAMA in test samples is determined directly from this standard curve