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Protein extraction from mature rice leaves for two‐dimensional gel electrophoresis and its application in proteome analysis
Author(s) -
Islam Nazrul,
Lonsdale M.,
Upadhyaya N. M.,
Higgins T. J.,
Hirano H.,
Akhurst R.
Publication year - 2004
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200300816
Subject(s) - proteome , extraction (chemistry) , chromatography , protein purification , two dimensional gel electrophoresis , gel electrophoresis , electrophoresis , sample preparation , proteomics , biology , chemistry , biochemistry , gene
Sample preparation is crucial for extraction and higher resolution of proteins by two‐dimensional gel electrophoresis (2‐DE). In this study, we present an efficient protocol to extract proteins from mature rice leaves by minimizing the presence of nonprotein contaminants and by maximizing contact between the sample and extraction buffer. A combination of chemical and physical processes remarkably improved protein extraction for 2‐DE. The efficiency of this protocol was demonstrated by comparison of the rice proteome at two developmental stages.