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Sample application by in‐gel rehydration improves the resolution of two‐dimensional electrophoresis with immobilized pH gradients in the first dimension
Author(s) -
Rabilloud Thierry,
Valette Cecyl,
Lawrence Jean Jacques
Publication year - 1994
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.11501501223
Subject(s) - isoelectric focusing , chromatography , immobilized ph gradient , resolution (logic) , sample (material) , sample preparation , chemistry , electrophoresis , analytical chemistry (journal) , dimension (graph theory) , materials science , computer science , mathematics , artificial intelligence , enzyme , biochemistry , pure mathematics
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.