Phospho-Smad4 (T277) and Total Smad4 ELISA Kit

Product details

Phospho-Smad4 (T277) and Total Smad4 ELISA Kit (ab279932) 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 Smad4 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-Smad4 and total Smad4. An anti-pan Smad4 antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Smad4 present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho Smad4 (T277) antibody is added to detect phosphorylated Smad4. In the remaining wells, biotinylated anti-pan-Smad4 antibody is used to detect pan Smad4. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin 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 Smad4 (T277) or pan Smad4 bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

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