Phospho-CREB (S133) and Total CREB ELISA Kit (ab279764) 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 CREB 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-CREB and total CREB. An anti-pan CREB antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and CREB present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho-CREB (S133) antibody is added to detect phosphorylated CREB. In the remaining wells, mouse anti-pan-CREB antibody is used to detect pan CREB. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-conjugated anti-mouse 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 CREB (S133) or pan CREB bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.