Akt1 S1 (Phospho-Thr246)Colorimetric Cell-Based ELISA Kit

Assay Genie offers a Cell-Based ELISA kit designed to detect phosphorylated AKT1S1 at threonine 246 directly in adherent cell samples. The colorimetric assay uses an indirect ELISA format, in which target-specific primary antibodies are captured and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition, with absorbance read at 450 nm. The kit is validated for Human, Mouse, and Rat reactivity, has a stated dynamic range of > 5000 Cells, and is described as a convenient, lysate-free, high throughput and sensitive format.

Beyond measuring relative levels of phosphorylated AKT1S1, the kit can be used to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on AKT1S1 phosphorylation. Because the Cell-Based ELISA is qualitative, a monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used to determine cell density, allowing absorbance values to be normalised to cell amounts.

AKT1S1 (Proline-rich AKT1 substrate 1) is a subunit of mTORC1 that negatively regulates mTOR activity in a manner dependent on its phosphorylation state and binding to 14-3-3 proteins, playing a role in cell growth, survival and metabolism. Database identifiers listed for the target include Gene ID 84335, UniProt ID Q96B36 and OMIM 610221, and the encoded protein has a molecular weight of 29,262 Da.