Enhanced Protein Extraction from Tobacco Roots for Proteomic Analysis
ANALYTICAL LETTERS(2015)
Abstract
Plant roots contain low protein concentrations and many interferences for protein extraction and two-dimensional electrophoresis analysis. Therefore, the extraction of high-quality protein from tobacco roots for proteomic analysis is a challenge. Three protein extraction methods (the trichloroacetic acid-acetone, phenol extraction, and trichloroacetic acid-acetone-phenol methods) for tobacco root proteins were compared using protein yields, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and two-dimensional electrophoresis. The trichloroacetic acid-acetone-phenol method provided a higher spot resolution (505 +/- 18 spots), the least streaking, and larger protein yields (2200 +/- 20 mu g/g fresh weight) on two-dimensional electrophoresis gels for tobacco roots, and hence is the most suitable method for the characterization of tobacco roots.
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Key words
Proteomic analysis,High-quality protein extraction,Two-dimensional electrophoresis,Tobacco roots
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