|
|||||
|
|
Two-dimensional maps in very acidic immobilized pH gradients |
|
| Authors: | Pranav K. Sinha E. K ttgen Marina St ffler-Meilicke Elisabetta Gianazza Pier Giorgio Righetti |
| Affiliation: | a Institute of Clinical Chemistry and Biochemistry, Freie Universität Berlin, Universitätsklinikum Rudolf Virchow, Berlin, F.R.G. b Department of Virology, Freie Universität Berlin, Hindenburg Damm, Berlin, F.R.G. c Faculty of Pharmacy and Dept. Biomedical Sciences and Technologies, University of Milan, Milan, Italy |
| Abstract: | Up to the present time it has been impossible to perform two-dimensional (2-D) separations in very acidic immobilized pH gradients (IPG), due to the lack of suitable buffering acrylamido derivatives to be incorporated into the polyacrylamide matrix. The advent of the pK 3.1 buffer (2-acrylamido glycolic acid; Righetti et al., J. Biochem. Biophys. Methods 16, 1988, 185–192) allowed the formulation of such acidic gradients. We report here separations in IPG pH 2.8–5.0 intervals of polypeptide chains from total lysates of rat intestinal and liver cells and 30S and 50S ribosomal proteins from Halobacterium marismortui. Conditions are given for highly reproducible first and second dimensions gels and for a proper silver staining of 2-D maps with practically no background deposition. |
| Keywords: | Immobilized pH gradient Isoelectric focusing Two-dimensional map |
| 本文献已被 ScienceDirect 等数据库收录！ | |
