Analysis of proteostasis during aging with western blot of detergent-soluble and insoluble protein fractions

STAR Protoc. 2021 Jun 23;2(3):100628. doi: 10.1016/j.xpro.2021.100628. eCollection 2021 Sep 17.

Abstract

Defects in protein quality control are the underlying cause of age-related diseases. The western blot analysis of detergent-soluble and insoluble protein fractions has proven useful in identifying interventions that regulate proteostasis. Here, we describe the protocol for such analyses in Drosophila tissues, mouse skeletal muscle, human organoids, and HEK293 cells. We describe key adaptations of this protocol and provide key information that will help modify this protocol for future studies in other tissues and disease models. For complete details on the use and execution of this protocol, please refer to Rai et al. (2021) and Hunt el al. (2021).

Keywords: Cell Biology; Developmental biology; Model Organisms; Molecular Biology; Organoids; Protein Biochemistry.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / methods*
  • Detergents / chemistry*
  • Electrophoresis, Polyacrylamide Gel
  • HEK293 Cells
  • Humans
  • Mice
  • Proteins / chemistry
  • Proteins / metabolism*
  • Proteostasis*
  • Solubility
  • Ubiquitination

Substances

  • Detergents
  • Proteins