This AMPK alpha (phospho Thr172) Cell Based ELISA Kit allows for the detection of AMPK alpha (phospho Thr172) and the effects that certain stimulation conditions have on AMPK alpha (phospho Thr172) expression in different cell lines. Qualitative determination of AMPK alpha (phospho Thr172) concentration is achieved by an indirect ELISA format. In essence, AMPK alpha (phospho Thr172) is captured by Anti-AMPK alpha (phospho Thr172) Antibodies while Dye 1-conjugated and Dye 2-conjugated secondary antibodies bind the Fc region of the primary antibody. Through this binding, the dye conjugated to the secondary antibody can emit light at a certain wavelength given proper excitation, hence allowing for a fluorometric detection method. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target RFU values. Anti-AMPK alpha Antibody is also included for normalization purposes. The RFU values obtained for the non-phosphorylated target can be used to normalize the RFU value for the phosphorylated target.