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

• Published in: Analytical biochemistry Vol. 421; no. 1; pp. 330 - 332
• Main Authors: Young, Clifford, Truman, Penelope
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-02-2012
• Subjects: 2-DE; Amino Acid Sequence; Animals; Cell Line, Tumor; desorption; digestion; Electrophoresis, Gel, Two-Dimensional - methods; gels; Guanidines; Horses; Humans; ionization; Mass spectrometry; Molecular Sequence Data; myoglobin; Myoglobin - chemistry; Myoglobin - isolation & purification; Neoplasm Proteins - chemistry; Neoplasm Proteins - isolation & purification; Peptide Mapping - methods; Peptide mass fingerprint; peptides; Phenols; Protein extraction; Proteins - chemistry; Proteins - isolation & purification; Proteomics; Proteomics - methods; RNA; sodium dodecyl sulfate; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; TRIzol; Trypsin; two-dimensional gel electrophoresis; Protein extraction; TRIzol; Peptide mass fingerprint; 2-DE; Mass spectrometry; Proteomics
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2011.10.045

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.