Extraction methods for analysis of citrus leaf proteins by two-dimensional gel electrophoresis.
General procedures for the extraction of Citrus leaf proteins for analysis by two-dimensional electrophoresis (2-DE) were developed through the evaluation and modification of existing methods. Among the methods evaluated, the best results were obtained when Tris-HCl, KCl, and phenol extractions were followed by precipitation with organic solvents to purify and concentrate the samples. The utility of the Tris-HCl extraction method was demonstrated on the leaves of six genetically different Citrus varieties. The methods described are versatile and result in high resolution 2-DE gels of Citrus leaf proteins.
Zukas
Audrius A
AA
Western Regional Research Center, Agricultural Research Service, United States Department of Agriculture, 800 Buchanan St., Albany, CA 94710, USA.
Breksa
Andrew P
AP
D005591:Chemical Fractionation; D002957:Citrus / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D019800:Phenol; D018515:Plant Leaves / Q000737:chemistry*; D010940:Plant Proteins / Q000032:analysis* / Q000302:isolation & purification*; D011189:Potassium Chloride; D040901:Proteomics; D014325:Tromethamine
D010940:Plant Proteins; D014325:Tromethamine; D019800:Phenol; D011189:Potassium Chloride