CrkII (Phospho-Tyr221)Colorimetric Cell-Based ELISA Kit

Assay Genie's CrkII (Phospho-Tyr221) Colorimetric Cell-Based ELISA Kit is designed for the accurate and sensitive detection of phosphorylated CRKII protein at tyrosine 221 in cell lysates. The cell-based assay is lysate-free, high throughput, and uses a colorimetric detection method read at 450 nm, with reactivity confirmed for human, mouse, and rat samples. The kit supports a dynamic range of > 5000 cells.

Qualitative determination of CrkII (Phospho-Tyr221) is achieved by an indirect ELISA format, in which CrkII (Phospho-Tyr221)-specific primary antibodies capture the target and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalizing target absorbance values, and Crystal Violet whole-cell staining is used to determine cell density so plating differences can be adjusted.

Intended for use with adherent cells, the kit can be used to measure relative levels of phosphorylated CrkII in cultured cells, as well as to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on CrkII phosphorylation. CRKII is an SH2-SH3-SH3 adaptor protein involved in cell growth, differentiation, and migration, and phosphorylation at tyrosine 221 is known to modulate its activity and downstream signaling.