Principle of Assay
Mouse TNF alpha ELISA Kit (A335185) is an Indirect ELISA for the quantitative measurement of mouse TNF alpha in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The 96-well microtiter plate has been pre-coated with TNF alpha antigen. The samples and standards are added to the wells and incubated. After washing with wash buffer, Biotinylated detection antibody is added and incubated. After washing with wash buffer, HRP-Streptavidin is then added and incubated. Following another wash step, TMB substrate is added to visualize the HRP enzymatic reaction. TMB is catalysed by HRP to produce a blue-coloured product that changes to yellow after the addition of the acidic stop solution. The density of yellow is proportional to the amount of TNF alpha sample captured on the plate. The concentration of TNF alpha can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.
