Far-western Blotting Detection of the Binding of Insulin Receptor Substrate to the Insulin Receptor
Jinghua Peng,
Balamurugan Ramatchandirin,
Alexia Pearah,
Ling He
Affiliations
Jinghua Peng
Institute of Liver Diseases, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai, ChinaDepartment of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
Balamurugan Ramatchandirin
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
Alexia Pearah
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
Ling He
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, Maryland, USADepartment of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA
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.
