HTRF® phospho-protein platform facilitates the dissection of signaling pathways involved in insulin resistance and metabolic disorders

Julie Vallaghé1, Nathalie Costy1, Vincent Morcillo1, Michael Mathieu1, Coby Carlson2, Julia Mathieu3, Stéphane Dalle3, Jean-François Tanti4 and Fabienne Charrier-Savournin1

Cisbio, IGF, CDI

Poster

Cisbio has developed a broad range of HTRF® (Homogeneous Time-Resolved Fluorescence) cell-based immunoassays to analyze the phosphorylation and/or the expression level of key intracellular signaling proteins. These homogeneous “add-and-read” assays based on TR-FRET are sensitive, accurate and most importantly, require low sample volumes. Here, we demonstrate that HTRF cell signaling assays are a straightforward and robust solution for the study of proteins involved in insulin signaling activation/inhibition. This poster shows how we analyzed the IRS1/2-AKT-GSK3β/mTOR/FoxO1 and AMPK/ACC metabolic signaling axes, as well as the IKK and JNK stress pathways, using various relevant cellular models such as rat pancreatic β-cells, mouse adipocytes, and human iPSC-derived hepatocytes.

Phosphorylated proteins