Two-dimensional electrophoresis of basic proteins with equilibrium isoelectric focusing in carrier ampholyte-pH gradients
- 1 January 1994
- journal article
- Published by Wiley in Electrophoresis
- Vol. 15 (1), 278-282
- https://doi.org/10.1002/elps.1150150147
Abstract
A modified procedure for the two‐dimensional electrophoretic analysis of basic polypeptides is described. This method uses isoelectric focusing with carrier ampholytes in the first dimension, and sodium dodecyl sulfate‐electrophoresis in the second dimension. Counteraction of the cathodic drift is achieved by glass tube treatment (silanization), electrolyte modification (use of weak bases and acids), protection of the catholyte from carbon dioxide, and the addition of glycerol to the gel mix. Better resolution and reproducibility are obtained than with nonequilibrium pH gradient electrophoresis, since quasi equilibrium focusing can be obtained.Keywords
This publication has 22 references indexed in Scilit:
- Approach to stationary two‐dimensional pattern: Influence of focusing time and Immobiline/carrier ampholytes concentrationsElectrophoresis, 1988
- Properties of thin‐rod immobilized pH gradientsElectrophoresis, 1988
- Removal of oligomers and n-mers from the immobiline chemicals for isoelectric focusingJournal of Chromatography A, 1987
- Isoelectric focusing and non-isoelectric precipitation of ferritin in immobilized pH gradients: An improved protocol overcoming protein-matrix interactionsElectrophoresis, 1987
- Protein precipitation induced by alkaline Immobilines for isoelectric focusing in immobilized pH gradients: Causes and remediesElectrophoresis, 1987
- High resolution two‐dimensional polyacrylamide gel electrophoresis of basic myeloid polypeptides: Use of immobilized pH gradients in the first dimensionElectrophoresis, 1986
- Factors affecting the range of pH gradients in the isoelectric focusing dimennsion of two-dimensional gel electrophoresis: The effects of reservoir electrolytes and loading proceduresElectrophoresis, 1985
- Immobilized pH gradients for isoelectric focusing: Interaction between histones and histone‐like proteins with the charged polyacrylamide matrixElectrophoresis, 1983
- Enhancement of resolution in two‐dimensional gel electrophoresis and simultaneous resolution of acidic and basic proteinsElectrophoresis, 1982
- High resolution two-dimensional electrophoresis of basic as well as acidic proteinsCell, 1977