Akt1 (Phospho-Ser473)Fluorometric Cell-Based ELISA Kit

From Assay Genie, this Phospho Cell-Based ELISA Kit is designed for the qualitative detection of Akt1 (Phospho-Ser473) in cultured cells using a fluorometric format. The assay employs an indirect ELISA approach in which target-specific primary antibodies capture Akt1 (Phospho-Ser473), while dye-conjugated secondary antibodies binding the Fc region of the primary antibody emit light at defined wavelengths for detection.

The kit is lysate-free and offers a high-throughput workflow suited to monitoring Akt1 phosphorylation and expression profiles, measuring relative amounts of phosphorylated Akt1 in cultured cells, and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on AKT1/AKT2/AKT3 phosphorylation. The reported dynamic range is > 5000 Cells, with reactivity confirmed for Human, Mouse, and Rat samples, and a storage/stability of 4°C/6 Months. Detection is performed at Ex/Em channels of 651/667 and 495/521, and associated database identifiers are Gene ID 207, UniProt ID P31749, OMIM # 164730, and Unigene Hs.525622.

For data normalization, the kit includes a monoclonal antibody specific for human GAPDH as an internal positive control, as well as an antibody against the non-phosphorylated counterpart of Akt1 (Phospho-Ser473), so that RFU values obtained for non-phosphorylated Akt1 can be used to normalize the RFU value for phosphorylated Akt1.