Phospho-BTK (Y551) and Total BTK ELISA Kit (ab279753) 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 BTK 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-BTK and total BTK. An anti-pan BTK antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and BTK present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho BTK (Y551) antibody is added to detect phosphorylated BTK (Y551). In the remaining wells, rabbit anti-pan-BTK antibody is used to detect pan BTK. 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 BTK (Y551) or pan BTK bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.