Phospho-Bad (S112) and Total Bad ELISA Kit (ab279746) is a very rapid, convenient, and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated Bad protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-Bad and total Bad. An anti-pan Bad antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Bad present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-Bad (S112) antibody is added to detect phosphorylated Bad. In the remaining wells, rabbit anti-pan-Bad antibody is used to detect pan Bad. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Bad (S112) or pan Bad bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.