Proteins isolated with TRIzol are compatible with two-dimensional electrophoresis and mass spectrometry analyses

Anal Biochem. 2012 Feb 1;421(1):330-2. doi: 10.1016/j.ab.2011.10.045. Epub 2011 Nov 4.

Abstract

TRIzol is used for RNA isolation but also permits protein recovery. We investigated whether proteins prepared with TRIzol were suitable for two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectrometry. Proteins from TRIzol-treated SH-SY5Y cells produced 2-DE spot patterns similar to those from an equivalent untreated sample. Subsequent identification of TRIzol-treated proteins using peptide mass fingerprinting was successful. TRIzol exposure altered neither the mass of myoglobin extracted from sodium dodecyl sulfate (SDS) gels nor the masses of myoglobin peptides produced by in-gel trypsin digestion. These findings suggest that proteins isolated with TRIzol remain amenable to proteomic analyses.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Cell Line, Tumor
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Guanidines*
  • Horses
  • Humans
  • Molecular Sequence Data
  • Myoglobin / chemistry
  • Myoglobin / isolation & purification
  • Neoplasm Proteins / chemistry
  • Neoplasm Proteins / isolation & purification
  • Peptide Mapping / methods
  • Phenols*
  • Proteins / chemistry
  • Proteins / isolation & purification*
  • Proteomics / methods
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*
  • Trypsin

Substances

  • Guanidines
  • Myoglobin
  • Neoplasm Proteins
  • Phenols
  • Proteins
  • trizol
  • Trypsin