Rearranged during transfection (RET or c-RET) is a receptor tyrosine kinase primarily expressed as two isoforms of 1072 & 1114 amino acids. RET activation by glial cell line-derived neurotrophic family ligands (GFL) leads to its dimerization and autophosphorylation on intracellular tyrosine residues. RET is required for the development of the nervous system and several other tissues. RET alterations have been detected in numerous human cancers. The development of highly selective and potent RET kinase inhibitors is therefore of great interest to treat RET-altered cancers.

The AlphaLISA SureFire Ultra Multiplex Human and Mouse Phospho-RET (panTyr)/Total RET Detection Kit is used to measure both phosphorylated and total levels of endogenous RET in cellular lysates in a multiplexing Alpha no-wash assay. No specially engineered cell lines are required. The 615 nm (Eu) signal corresponds to the phosphorylated RET analysis, and the 545 nm (Tb) signal corresponds to the total RET analysis. This kit has been formulated to provide superior signal:background assay windows and to perform without interference in the presence of extraneous antibodies, making it amenable to the study of therapeutic and blocking antibodies.

• No-wash steps, no separation steps
• Multiplexing assay
• Sensitive detection
• No interference from exogenous antibodies
• Results in less than 3 hours

In this assay, the Alpha 615 Acceptor bead is coated with the CaptSure^™ antibody, which binds the CaptSure-tagged anti-phospho target antibody. The Alpha 545 Acceptor bead is coated with the CaptSure2™ agent, which binds the CaptSure2 tagged anti-total target protein antibody. The Alpha Donor bead binds the biotinylated anti-total target protein antibody.