Protein solubilization: attend to the choice of lysis buffer

Methods Mol Biol. 2012:869:37-47. doi: 10.1007/978-1-61779-821-4_4.

Abstract

The efficient extraction of proteins of interest from cells and tissues is not always straightforward. In this process, the use of the optimal lysis buffer for protein solubilization should be considered. Here we demonstrate the use of a urea/thiourea lysis buffer, based on O'Farrell's buffer, and compare its effectiveness for solubilization of proteins from smooth muscle with the often utilized RIPA lysis buffer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western / methods
  • Buffers
  • Deoxycholic Acid / chemistry
  • Detergents / chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Muscle Proteins / chemistry
  • Muscle Proteins / isolation & purification*
  • Muscle, Smooth / chemistry
  • Muscle, Smooth / cytology
  • Octoxynol / chemistry
  • Protein Denaturation
  • Sodium Dodecyl Sulfate / chemistry
  • Solubility
  • Surface-Active Agents / chemistry
  • Thiourea / chemistry
  • Urea / chemistry

Substances

  • Buffers
  • Detergents
  • Muscle Proteins
  • Surface-Active Agents
  • Deoxycholic Acid
  • Sodium Dodecyl Sulfate
  • Urea
  • Octoxynol
  • Thiourea