AKT1/3 (Phospho-Tyr437/434)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the measurement of AKT1/3 (Phospho-Tyr437/434) phosphorylation in cell lysates and tissue samples. The kit is reactive with Human, Mouse, and Rat samples, has a dynamic range of > 5000 Cells, and uses a Colorimetric detection method at 450 nm within a Cell-Based ELISA format.

The lysate-free, high-throughput assay detects AKT1/3 protein phosphorylation and expression profiles in cells via an indirect ELISA format. Target-specific primary antibodies capture the protein, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyze a colorimetric reaction upon substrate addition. This enables measurement of the relative amounts of phosphorylated AKT1/3 in cultured cells and screening of effects from various treatments, inhibitors such as siRNA or chemicals, or activators on AKT1/3 phosphorylation.

For data normalization, a monoclonal antibody specific for human GAPDH is included as an internal positive control. Following the colorimetric measurement, the Crystal Violet whole-cell staining method may be used to determine cell density, allowing absorbance values to be normalized to cell amounts. AKT1 is a key signaling protein involved in cell growth, survival, and metabolism, and phosphorylation at Tyr437/434 is known to regulate its activity. Dysregulation of AKT1 phosphorylation has been implicated in cancer, diabetes, and neurological disorders, making it a target of interest for biomarker discovery and therapeutic development research.