Western Blotting and Co-immunoprecipitation of Endogenous STIM/ORAI and Protein Partners

Methods Mol Biol. 2018;1843:107-113. doi: 10.1007/978-1-4939-8704-7_9.

Abstract

The characterization of protein-protein interactions through methods such as co-immunoprecipitation, followed by Western blot analysis, is a crucial step in the understanding of protein functions and the biology of the cell. Since the discovery of ORAI and STIM proteins as component of store-operated channel (SOC), overexpressing systems have been used to demonstrate how ORAI and STIM can associate with physiological and pathological conditions. Here we describe a protocol allowing endogenous studies.

Keywords: Co-immunoprecipitation; Endogenous; ORAI; STIM; Western blot.

MeSH terms

  • Blotting, Western* / methods
  • Calcium / metabolism
  • Calcium Channels / metabolism
  • Carrier Proteins / metabolism*
  • Cell Line, Tumor
  • Humans
  • Immunoprecipitation* / methods
  • Protein Binding
  • Protein Interaction Mapping* / methods
  • Stromal Interaction Molecules / metabolism*

Substances

  • Calcium Channels
  • Carrier Proteins
  • Stromal Interaction Molecules
  • Calcium