Description
1. Cells are seeded onto the bottom of each well. The cells are quenched, fixed and the well is blocked.
2. Primary antibodies specific for the target antigen are added and allowed to bind to their respective epitopes.
3. HRP-conjugated secondary antibodies specific for the primary antibody are added and allowed to bind to their respective epitopes.
4. TMB substrateis converted to the blue TMB diimine via the HRP enzyme.Upon addition of acid,the reaction terminates and the wells can be read at 450nm