Resolubilization of TCA precipitated plant proteins for 2-D electrophoresis

Electrophoresis. 2011 Mar;32(6-7):696-8. doi: 10.1002/elps.201000557. Epub 2011 Feb 21.

Abstract

In this short communication, a novel protocol for resolubilization of TCA-precipitated plant proteins for 2-DE is described. Guanidine hydrochloride (Gdn-HCl) is used as an intermediate for protein solubilization in which proteins are reduced and alkylated with tributylphosphane (TBP) and 2-vinylpyridine (2-VP). The blocking of -SH groups at Cys residues can greatly improve the solubility of TCA-precipitated proteins and obtain more high-quality protein spots in the 2-DE gel. This protocol is compatible with silver stain and MS identification.

MeSH terms

  • Alkylation
  • Arachis / chemistry
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Guanidine
  • Phosphines / chemistry
  • Plant Leaves / chemistry
  • Plant Proteins / chemistry*
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism
  • Pyridines / chemistry
  • Reproducibility of Results
  • Silver Staining
  • Solubility
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Trichloroacetic Acid / chemistry*

Substances

  • Phosphines
  • Plant Proteins
  • Pyridines
  • tri-n-butylphosphine
  • Trichloroacetic Acid
  • 2-vinylpyridine
  • Guanidine