AMPKalpha (Phospho-Thr172)Fluorometric Cell-Based ELISA Kit

This cell-based ELISA kit from Assay Genie is designed for monitoring AMPKalpha phosphorylation at Thr172 in cultured cells. It is a lysate-free, high-throughput, fluorometric assay that uses an indirect ELISA format: a phospho-Thr172-specific primary antibody captures the target, and Dye 1- and Dye 2-conjugated secondary antibodies bound to the Fc region generate a fluorescent signal at specific excitation/emission wavelengths. Plates are read at Ex/Em 651/667 nm (Dye 1) and 495/521 nm (Dye 2).

The kit is reactive with Human, Mouse, and Rat samples, with a stated dynamic range of > 5000 cells. Storage is 4 °C for 6 months. Two normalization reagents are provided: an anti-GAPDH monoclonal antibody serves as an internal positive control for normalizing target RFU values, and an antibody against the non-phosphorylated AMPKalpha counterpart is included so that phosphorylated RFU values can be normalized to total AMPKalpha signal. The associated NCBI gene symbols are AAPK1/AAPK2 (UniProt Q13131/P54646; Gene IDs 5562/5563).

Typical uses include measuring relative amounts of phosphorylated AMPKalpha in cultured cells and screening the effects of treatments, inhibitors (e.g., siRNA or chemicals), or activators on AMPKalpha phosphorylation.