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Improvement of the two‐dimensional gel electrophoresis analysis for the proteome study of Halobacterium salinarum
Author(s) -
Cho ChangWon,
Lee SoHee,
Choi Jiyeon,
Park SooJin,
Ha DongJin,
Kim HyoJeong,
Kim ChanWha
Publication year - 2003
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.200300525
Subject(s) - halobacterium salinarum , proteome , gel electrophoresis , two dimensional gel electrophoresis , proteomics , chemistry , electrophoresis , polyacrylamide gel electrophoresis , biology , chromatography , biochemistry , archaea , gene , enzyme
Inherent problems exist in the use of two‐dimensional gel electrophoresis (2‐DE) for sample preparation and separation of proteins from Halobacterium salinarum . In particular, proteins from cells grown in 25% NaCl are difficult to resolve by 2‐DE due to the abundance of salt. To remove salts, a 3 kDa molecular weight cut‐off column was used. When soluble proteins were separated by 2‐DE, most of the proteins were concentrated in the acidic range. For separation of proteins in the pH 3–6 range, ultrazoom immobilized pH gradient strips were used. In addition, sample separation using a IPGphor/Multiphor combined system was a more effective method for the proteome analysis of acidic proteins than using IPGphor for the isoelectric focusing step.