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Two-dimensional maps in very acidic immobilized pH gradients.
- Source :
-
Journal of biochemical and biophysical methods [J Biochem Biophys Methods] 1990 Apr-May; Vol. 20 (4), pp. 345-52. - Publication Year :
- 1990
-
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 dimension gels and for a proper silver staining of 2-D maps with practically no background deposition.
Details
- Language :
- English
- ISSN :
- 0165-022X
- Volume :
- 20
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Journal of biochemical and biophysical methods
- Publication Type :
- Academic Journal
- Accession number :
- 2365950
- Full Text :
- https://doi.org/10.1016/0165-022x(90)90096-u