title: Universal sample preparation method integrating trichloroacetic acid/acetone precipitation with phenol extraction for crop proteomic analysis.
abstract: Crop plants contain large amounts of secondary compounds that interfere with protein extraction and gel-based proteomic analysis. Thus, a protein extraction protocol that can be easily applied to various crop materials with minimal optimization is essential. Here we describe a universal protocol for total protein extraction involving trichloroacetic acid (TCA)/acetone precipitation followed by SDS and phenol extraction. Through SDS extraction, the proteins precipitated by the TCA/acetone treatment can be fully resolubilized and then further purified by phenol extraction. This protocol combines TCA/acetone precipitation, which aggressively removes nonprotein compounds, and phenol extraction, which selectively dissolves proteins, resulting in effective purification of proteins from crop tissues. This protocol can also produce high-quality protein preparations from various recalcitrant tissues, and therefore it has a wide range of applications in crop proteomic analysis. Designed to run on a small scale, this protocol can be completed within 5 h.
authors.affiliation: Synergetic Innovation Center of Henan Grain Crops, Key Laboratory of Physiological Ecology and Genetic Improvement of Food Crops in Henan Province, College of Life Sciences, Henan Agricultural University, Zhengzhou, China.
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Wu
authors.affiliation: Synergetic Innovation Center of Henan Grain Crops, Key Laboratory of Physiological Ecology and Genetic Improvement of Food Crops in Henan Province, College of Life Sciences, Henan Agricultural University, Zhengzhou, China.
authors.forename: Erhui
authors.initials: E
authors.lastname: Xiong
authors.affiliation: Synergetic Innovation Center of Henan Grain Crops, Key Laboratory of Physiological Ecology and Genetic Improvement of Food Crops in Henan Province, College of Life Sciences, Henan Agricultural University, Zhengzhou, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Scali
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy.
authors.forename: Mauro
authors.initials: M
authors.lastname: Cresti
mesh_terms: D000096:Acetone / Q000737:chemistry*; D053000:Analytic Sample Preparation Methods / Q000379:methods*; D018556:Crops, Agricultural / Q000235:genetics* / Q000378:metabolism; D010636:Phenols; D020543:Proteome / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D012967:Sodium Dodecyl Sulfate; D014238:Trichloroacetic Acid / Q000737:chemistry*
chemical_list: D010636:Phenols; D020543:Proteome; D000096:Acetone; D012967:Sodium Dodecyl Sulfate; D014238:Trichloroacetic Acid