STAT5A/B (Phospho-Ser731)Colorimetric Cell-Based ELISA Kit

Assay Genie's in-cell ELISA kit is designed for the precise measurement of STAT5A/B phosphorylated at Ser731 in cultured cells. The assay employs a colorimetric detection method with absorbance read at 450 nm in an indirect ELISA format, where phospho-specific primary antibodies capture the target and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition. The kit is lysate-free, supports high-throughput workflows, and is intended for adherent cell samples, with reactivity against human, mouse, and rat targets.

STAT5A/B is a transcription factor involved in fundamental cellular processes including proliferation, differentiation, and survival, and its activation through phosphorylation at Ser731 is a key step in signal transduction. The kit can be used to measure relative levels of phosphorylated STAT5A/B in cultured cells and to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on STAT5A/B phosphorylation. A monoclonal antibody specific for human GAPDH is included as an internal positive control to normalize target absorbance values, and a Crystal Violet whole-cell staining step allows determination of cell density, so absorbance values can be corrected for plating differences.

The dynamic range of the assay is > 5000 cells, and the sub-type is phospho-specific, targeting STAT5A/STAT5B.