Universal sample preparation method integrating trichloroacetic acid/acetone precipitation with phenol extraction for crop proteomic analysis View Full Text


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Article Info

DATE

2014-01-16

AUTHORS

Xiaolin Wu, Erhui Xiong, Wei Wang, Monica Scali, Mauro Cresti

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. More... »

PAGES

362-374

References to SciGraph publications

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  • 2010-12-28. Proteomic identification of differentially expressed proteins in mature and germinated maize pollen in ACTA PHYSIOLOGIAE PLANTARUM
  • 2010-06-19. Optimization of Protein Extraction from Hypericum perforatum Tissues and Immunoblotting Detection of Hyp-1 at Different Stages of Leaf Development in MOLECULAR BIOTECHNOLOGY
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  • 2009-11-13. Changes in the accumulation of α- and β-tubulin during bud development in Vitis vinifera L. in PLANTA
  • 2010-05-22. Characterization of Small Heat Shock Proteins Associated with Maize Tolerance to Combined Drought and Heat Stress in JOURNAL OF PLANT GROWTH REGULATION
  • 2010-05-22. Proteomic analysis of leaves from a diploid cybrid produced by protoplast fusion between Satsuma mandarin and pummelo in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
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  • 2010-06-05. Proteomic analysis of seed viability in maize in ACTA PHYSIOLOGIAE PLANTARUM
  • 2006-07-13. Sample extraction techniques for enhanced proteomic analysis of plant tissues in NATURE PROTOCOLS
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nprot.2014.022

    DOI

    http://dx.doi.org/10.1038/nprot.2014.022

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1030450829

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/24434803


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