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Horizontal two‐dimensional electrophoresis with immobilized pH gradients in the first dimension in the presence of nonionic detergent
Author(s) -
Görg Angelika,
Postel Wilhelm,
Weser Johann,
Günther Siegfried,
Strahler John R.,
Hanash Samir M.,
Somerlot Luke
Publication year - 1987
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150080110
Subject(s) - immobilized ph gradient , electrophoresis , chromatography , two dimensional gel electrophoresis , polyacrylamide gel electrophoresis , chemistry , gel electrophoresis , polyacrylamide , temperature gradient gel electrophoresis , gel electrophoresis of proteins , urea , biochemistry , isoelectric focusing , enzyme , polymer chemistry , proteomics , 16s ribosomal rna , gene
Horizontal two‐dimensional (2‐D) electrophoresis with immobilized pH gradients (IPG) in the first dimension, described for soluble proteins in Electrophoresis 1985, 6, 599–604, has been extended to an analysis of complex protein mixtures, such as leukemia cell proteins or bean proteins, in the presence of nonionic detergents. By optimizing the ratio of gel volumes for the first and second dimensional separation, and by decreasing the concentration of Nonidet P‐40 in the IPG gel, undistorted protein patterns in the 2‐D gel are obtained. Horizontal and vertical streaking are considerably diminished by optimized reswelling conditions of the dry IPG strips in presence of both urea and detergent. Resolution is improved and a reduced background is obtained when overcrowded gel areas are spread by flattening the pH gradient (in the first dimension) and the polyacrylamide gradient (in the second dimension).