Sample application by in‐gel rehydration improves the resolution of two‐dimensional electrophoresis with immobilized pH gradients in the first dimension
- 1 January 1994
- journal article
- research article
- Published by Wiley in Electrophoresis
- Vol. 15 (1) , 1552-1558
- https://doi.org/10.1002/elps.11501501223
Abstract
We describe a modification in the sample application mode for isoelectric focusing with immobilized pH gradients. Instead of being applied at the surface of the gel in a sample cup, the sample is introduced into the gel during the immobilized pH gradient strip rehydration step. This modification implies the use of low percentage gels (below 3.5%T) and specially designed, but simple, rehydration chambers. The main advantages are a uniform resolution without side effects and the possibility of handling large sample volumes (500 μL for a standard 3 × 160 × 0.5 mm strip), allowing micropreparative work (milligram samples) with a simple experimental design.Keywords
This publication has 27 references indexed in Scilit:
- Micropreparative two‐dimensional electrophoresis allowing the separation of samples containing milligram amounts of proteinsElectrophoresis, 1993
- The focusing positions of polypeptides in immobilized pH gradients can be predicted from their amino acid sequencesElectrophoresis, 1993
- A comparison between low background silver diammine and silver nitrate protein stainsElectrophoresis, 1992
- Immobilized pH 2.5–11 gradients for two‐dimensional electrophoresisElectrophoresis, 1992
- The Immobiline family: From “vacuum” to “plenum” chemistryElectrophoresis, 1992
- A new multiphasic buffer system for sodium dodecyl sulfate‐polyacrylamide gel electrophoresis of proteins and peptides with molecular masses 100 000–1000, and their detection with picomolar sensitivityElectrophoresis, 1991
- A reliable two‐dimensional gel electrophoresis procedure for separating neural proteinsElectrophoresis, 1991
- Two‐dimensional electrophoresis. The current state of two‐dimensional electrophoresis with immobilized pH gradientsElectrophoresis, 1988
- Horizontal two‐dimensional electrophoresis with immobilized pH gradients in the first dimension in the presence of nonionic detergentElectrophoresis, 1987
- 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