CRYAB (Phospho-Ser45)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the qualitative measurement of CRYAB phosphorylated at Ser45 directly in cultured cells. The assay uses an indirect ELISA format in which a phospho-specific primary antibody captures CRYAB (Phospho-Ser45) and an HRP-conjugated secondary antibody enables colorimetric detection at 450 nm. It is described as a convenient, lysate-free, high-throughput and sensitive In-Cell ELISA, suitable for adherent cells and for screening the effects of treatments, inhibitors (such as siRNA or chemicals) or activators on CRYAB phosphorylation.

The kit is reactive with human, mouse and rat samples and has a stated dynamic range of > 5000 cells. Because the Cell-Based ELISA is qualitative, two normalization methods are provided: a monoclonal antibody specific for human GAPDH is included as an internal positive control, and a Crystal Violet whole-cell staining step can be used to determine cell density and adjust for plating differences. CRYAB (alpha-B crystallin) is a member of the small heat shock protein (HSP20) family and is implicated in the cellular response to stress, with relevance to cardiovascular disorders and neurodegenerative conditions. The workflow follows a standard cell-based ELISA procedure: cells are fixed, quenched, blocked, and incubated with primary antibody overnight at 4 °C, followed by washing, secondary antibody incubation, and colorimetric substrate development.