Akt1 (Phospho-Ser246)Colorimetric Cell-Based ELISA Kit

Manufactured by Assay Genie, this cell-based ELISA is designed to detect phosphorylated AKT1 at serine 246 in cultured cells, supporting research into signaling pathways relevant to cancer, diabetes, and other diseases. The assay uses an indirect colorimetric format with HRP-conjugated secondary antibodies, yielding a readout at 450 nm, and is reactive with human, mouse, and rat samples. The dynamic range is listed as > 5000 Cells.

Because the kit is lysate-free, cells are fixed and processed in-plate, with a monoclonal anti-GAPDH antibody supplied as an internal positive control for normalization and Crystal Violet whole-cell staining used to correct absorbance values for cell density. The qualitative indirect ELISA workflow captures the target with phospho-specific primary antibodies, after which HRP-conjugated secondaries catalyze a colorimetric reaction on substrate addition. Plates are provided in a 96-well microplate format suitable for adherent cell work and for screening the effects of treatments, inhibitors (such as siRNA or chemicals), and activators on AKT1 phosphorylation.

AKT1, also known as protein kinase B, is documented in UniProt (entry P31749; Gene ID 207; chromosomal location 14q32.33) as an oncogenic AGC kinase that requires dual phosphorylation for activation, with T308 phosphorylated by PDK1 in the PI3 kinase pathway and S473 phosphorylated by mTOR in the mTORC2 pathway. The same entry notes that AKT1 is hyperactive or overexpressed in breast, prostate, lung, pancreatic, liver, ovarian, and colorectal cancers and that it acts on over 160 known protein substrates, making the phospho-Ser246 site a useful marker for studies of cell survival, metabolism, and signaling pathway activity.