Abstract
Polyacrylamide gel electrophoresis (PAGE) is a widely used technique for separating proteins from complex plant samples. Prior to the analysis, proteins must be extracted from plant tissues, which are rather complex than other types of biological material. Different protocols have been applied depending on the protein source, such as seeds, pollen, leaves, roots, and flowers. Total protein amounts must also be determined before conducting gel electrophoresis. The most common methodologies include PAGE under native or denaturing conditions. Both procedures are used consequently for protein identification and characterization via mass spectrometry. Additionally, various staining procedures are available to visualize protein bands in the gel, facilitating the software-based digital evaluation of the gel through image acquisition.
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References
Garfin DE (1990) One-dimensional gel electrophoresis. Methods Enzymol 182:425–441. https://doi.org/10.1016/0076-6879(90)82035-z. Bios Scientific, Oxford
Hames BD, Rickwood D (1990) Gel electrophoresis of proteins: a practical approach, vol 19. IRL Press at Oxford University Press, Oxford, p 39
Dunn MJ (1993) Gel electrophoresis of proteins. Bios Scientific, Oxford, p 176
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685. https://doi.org/10.1038/227680a0
Marzban G, Herndl A, Maghuly F et al (2008) Map** of fruit allergens by 2D electrophoresis and immunodetection. Expert Rev Proteomics 5:61–75. https://doi.org/10.1586/14789450.5.1.61
Wu FS, Wang MY (1984) Extraction of proteins for sodium dodecyl sulfate-polyacrylamide gel electrophoresis from protease-rich plant tissues. Anal Biochem 13:100–103. https://doi.org/10.1016/0003-2697(84)90394-4
Faurobert M, Pelpoir E, Chaïb J (2007) Phenol extraction of proteins for proteomic studies of recalcitrant plant tissues. Methods Mol Biol 355:9–14. https://doi.org/10.1385/1-59745-227-0:9
Wang W, Tai F, Chen S (2008) Optimizing protein extraction from plant tissues for enhanced proteomics analysis. J Sep Sci 31:2032–2039. https://doi.org/10.1002/jssc.200800087
Orr AR, Wagner BA, Howard CT et al (1988) Assay of plant proteins with bicinchoninic acid for high resolution two-dimensional polyacrylamide gel electrophoresis. Plant Cell Rep 7:598–601. https://doi.org/10.1007/BF00272767
Cawood AH, Potter U, Dickinson HG (1978) An evaluation of Coomassie Brilliant Blue as a stain for quantitative microdensitometry of protein in section. J Histochem Cytochem 26:645–650
Chevallet M, Luche S, Rabilloud T (2006) Silver staining of proteins in polyacrylamide gels. Nat Protoc 1:1852–1858. https://doi.org/10.1038/nprot.2006.288
Dowsey AW, Morris JS, Gutstein HB, Yang GZ (2010) Informatics and statistics for analyzing 2-d gel electrophoresis images. Methods Mol Biol 604:239–255. https://doi.org/10.1007/978-1-60761-444-9_16
West MJ, Webb HM, Sinclair AJ et al (2004) Biophysical and mutational analysis of the putative bZIP domain of Epstein-Barr virus EBNA 3C. J Virol 78:9431–9445. https://doi.org/10.1128/JVI.78.17.9431-9445.2004
Bartsch H, Arndt C, Koristka S et al (2012) Silver staining techniques of polyacrylamide gels. In: Kurien B, Scofield R (eds) Protein electrophoresis. Methods in molecular biology, vol 869. Humana Press, Totowa. https://doi.org/10.1007/978-1-61779-821-4_42
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Marzban, G., Tesei, D. (2024). One-Dimensional Acrylamide Gel Electrophoresis for Analysis of Plant Samples. In: Maghuly, F. (eds) Plant Functional Genomics. Methods in Molecular Biology, vol 2787. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3778-4_18
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DOI: https://doi.org/10.1007/978-1-0716-3778-4_18
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