STAT6 (Phospho-Tyr641)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the sensitive and specific detection of phosphorylated STAT6 at tyrosine 641. It is described as a convenient, lysate-free, high-throughput assay that uses an indirect ELISA format: STAT6 (Phospho-Tyr641) is captured by a phospho-specific primary antibody, while HRP-conjugated secondary antibodies bind the Fc region of that primary antibody and catalyze a colorimetric reaction upon substrate addition. The format allows qualitative determination of STAT6 (Phospho-Tyr641) concentration in cultured cells, supporting measurement of relative phosphorylation and screening of treatments, inhibitors such as siRNA, and activators.

The kit shows reactivity with Human, Mouse and Rat samples, with a stated dynamic range of > 5000 Cells and a colorimetric readout at 450 nm. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values, and the Crystal Violet whole-cell staining method can be used to determine cell density and adjust for plating differences. Adherent cells are supported as a sample type, and the workflow is built around seeded cells in microplate wells followed by fixation, blocking, primary and secondary antibody incubations, substrate addition and an immediate OD read at 450 nm.

Phosphorylation of STAT6 at tyrosine 641 is described as a critical event in the activation of the STAT6 signaling pathway, which regulates gene expression in response to interleukin-4 and interleukin-13 stimulation. Dysregulation of this pathway has been implicated in diseases such as asthma, allergy and autoimmune disorders, and STAT6 itself is a transcription factor of the STAT family with a central role in IL4-mediated biological responses, including the anti-apoptotic activity of IL4 via induction of BCL2L1/BCL-X(L) and involvement in T helper 2 cell differentiation and immunoglobulin class switching. The kit is positioned within the Epigenetics and Nuclear Signaling research area for studies of signal transduction, immune responses, inflammation and cell growth.