EnzyFluo™ Human/Mouse AKT1(S473) Phosphorylation ELISA Kit

• New and improved. Total assay time reduced from the standard 9 hours to 5 hours.
• Simple and convenient. Cells are directly cultured in 96-well plates. No cell lysis necessary.
• Accurate. Protein phosphorylation is normalized to total cellular protein in the same well, greatly minimizing well-to-well variations.

PROTEIN PHOSPHORYLATION plays key roles in regulating protein activity within cells. A large number of protein kinases and phosphatases are known to be involved in a variety of signal transduction pathways. A rapid assay capable of monitoring such signal transduction pathways, target protein phosphorylation status and drug effects is highly desirable in life science research and drug discovery.

AKT1 is a serine-threonine protein kinase that is activated through phosphatidylinositol 3-kinase by platelet-derived growth factor. In the developing nervous system AKT is a critical mediator of growth factorinduced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the AKT1. BioAssay Systems' AKT1(S473) Phosphorylation ELISA Kit provides a rapid and sensitive assay for measuring the phosphorylated protein pAKT1(S473) in cultured cells and normalizes the level of phosphorylation to the total protein content in the same well. This simple and efficient assay eliminates the need for cell lysate preparation and can be used to study the underlying signal pathway and effects of inhibitors, siRNA or activators on the pathway. In this assay, cells grown in a 96-well plate are fixed and permeabilized. The phosphorylation of AKT1(S473) is measured using a specific primary antibody followed by a HRP-conjugated secondary antibody with a fluorogenic substrate, and a fluorescent reagent that measures the total protein in the same well.