Use of Native-PAGE for the Identification of Epichaperomes in Cell Lines

Abstract

Epichaperomes are disease-associated pathologic scaffolds, composed of tightly bound chaperones, co-chaperones, and other factors. They mediate anomalous protein-protein interactions inside cells, which aberrantly affects the function of protein networks, and in turn, cellular phenotypes. Epichaperome study necessitates the implementation of methods that retain these protein complexes in their native cellular states for analysis. Here we describe a protocol for detection and composition analysis of epichaperomes in cell homogenates through native polyacrylamide gel electrophoresis.

Keywords: Chaperones; Epichaperomes; HSP70; Heat shock protein 90 (HSP90); High-order assemblies; Immunoblot; Multimolecular protein complexes; Native polyacrylamide gel electrophoresis (native-PAGE); Non-denaturing gel; Oligomers.