Akt1 (Phospho-Tyr326)Colorimetric Cell-Based ELISA Kit

This Colorimetric Cell-Based ELISA Kit from Assay Genie is designed for the qualitative detection of phosphorylated AKT1 at tyrosine 326 in cultured cells. It is a lysate-free, high-throughput and sensitive assay that measures relative levels of phosphorylated Akt1 and supports screening of treatments such as siRNA, chemicals or other activators and inhibitors for their effects on Akt1 phosphorylation. The kit shows reactivity with Human, Mouse and Rat samples and uses a colorimetric detection method at 450 nm, with a dynamic range of > 5000 Cells.

An indirect ELISA format is employed, in which target-specific primary antibodies capture the phosphorylated Akt1, and HRP-conjugated secondary antibodies catalyse a colour reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalisation, and a Crystal Violet whole-cell staining step can be used to determine cell density and correct for plating differences. The target corresponds to AKT1 (Gene ID: 207, UniProt ID: P31749), a serine/threonine kinase implicated in cell survival, proliferation and metabolism and dysregulated in diseases such as cancer, diabetes and inflammatory disorders.