Improvement of the two-dimensional gel electrophoresis analysis for the proteome study of Halobacterium salinarum.
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.
Cho
Chang-Won
CW
Graduate School of Life Sciences and Biotechnology, Korea University, 1, 5-ka Anam-dong, Sungbuk-ku, Seoul, Korea.
Lee
So-Hee
SH
Choi
Jiyeon
J
Park
Soo-Jin
SJ
Ha
Dong-Jin
DJ
Kim
Hyo-Jeong
HJ
Kim
Chan-Wha
CW
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
D020543:Proteome