title: Isolation of Oat (Avena sativa L.) Total Proteins and Their Prolamin Fractions for 2D Electrophoresis.
abstract: Appropriate sample preparation is essential to obtaining good results of two-dimensional gel electrophoresis (2-DE). For various reasons (particularly phenolic compounds, proteolytic enzymes, and cell-wall mucilages) the extraction of proteins from plant material, among them oat proteins, is difficult. During isolation all soluble substances that may interfere with the analysis (especially isoelectric focusing) are removed, and proteins of interest are separated from the remains. However, the applied procedure of isolation cannot be too extensive, because additional stages cause loss of the proteins.In this chapter, we describe a simple procedure for the isolation of oat total proteins and their prolamin fractions prior to 2-DE, without necessity of considerable purification. It can be used for oat protein fractionation, measurement of oat protein concentration, and their 2-DE analysis, with particular reference to prolamin fractions. The presented routine includes modified methods of plant seed proteins extraction and sequential Osborne extraction, based on oat protein solubility differences.
authors.affiliation: Chair of Food Biochemistry, University of Warmia and Mazury in Olsztyn, Plac Cieszyński 1, 10-726, Olsztyn, Poland. dota@uwm.edu.pl.
authors.forename: Dorota
authors.initials: D
authors.lastname: Nałęcz
authors.affiliation: Chair of Food Biochemistry, University of Warmia and Mazury in Olsztyn, Plac Cieszyński 1, 10-726, Olsztyn, Poland.
authors.forename: Marta
authors.initials: M
authors.lastname: Dziuba
authors.affiliation: Chair of Food Biochemistry, University of Warmia and Mazury in Olsztyn, Plac Cieszyński 1, 10-726, Olsztyn, Poland.
authors.forename: Iwona
authors.initials: I
authors.lastname: Szerszunowicz
mesh_terms: D018554:Avena / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D010936:Plant Extracts / Q000032:analysis / Q000737:chemistry; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism*; D055315:Prolamins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D020543:Proteome; D040901:Proteomics / Q000379:methods
chemical_list: D010936:Plant Extracts; D010940:Plant Proteins; D055315:Prolamins; D020543:Proteome
keywords: Avenins; Determination of protein concentration; Ethanol extraction of prolamins; Isolation; Oat proteins; Sample preparation for 2-DE