Far-western Blotting Detection of the Binding of Insulin Receptor Substrate to the Insulin Receptor

Jinghua Peng; Balamurugan Ramatchandirin; Alexia Pearah; Ling He

doi:10.21769/BioProtoc.4619

Far-western Blotting Detection of the Binding of Insulin Receptor Substrate to the Insulin Receptor

Bio Protoc. 2023 Feb 20;13(4):e4619. doi: 10.21769/BioProtoc.4619.

Authors

Jinghua Peng^{1

2}, Balamurugan Ramatchandirin², Alexia Pearah², Ling He^{2

3}

Affiliations

¹ Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, China.
² Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
³ Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.

Abstract

Far-western blotting, derived from the western blot, has been used to detect interactions between proteins in vitro, such as receptor-ligand interactions. The insulin signaling pathway plays a critical role in the regulation of both metabolism and cell growth. The binding of the insulin receptor substrate (IRS) to the insulin receptor is essential for the propagation of downstream signaling after the activation of the insulin receptor by insulin. Here, we describe a step-by-step far-western blotting protocol for determining the binding of IRS to the insulin receptor.

Keywords: Insulin receptor; Insulin receptor substrate; Post-translational modification; Protein–protein binding; Signal transduction.

Abstract

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