title: Improvement of the two-dimensional gel electrophoresis analysis for the proteome study of Halobacterium salinarum.
abstract: 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.
authors.affiliation: Graduate School of Life Sciences and Biotechnology, Korea University, 1, 5-ka Anam-dong, Sungbuk-ku, Seoul, Korea.
authors.forename: Chang-Won
authors.initials: CW
authors.lastname: Cho
authors.forename: So-Hee
authors.initials: SH
authors.lastname: Lee
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Choi
authors.forename: Soo-Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Dong-Jin
authors.initials: DJ
authors.lastname: Ha
authors.forename: Hyo-Jeong
authors.initials: HJ
authors.lastname: Kim
authors.forename: Chan-Wha
authors.initials: CW
authors.lastname: Kim
mesh_terms: D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D019617:Halobacterium salinarum / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D020543:Proteome