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Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels
Published on: August 22, 2007
Sample application by in-gel rehydration improves the resolution of two-dimensional electrophoresis with immobilized
T Rabilloud1, C Valette, J J Lawrence
1CEA-Laboratoire de Biologie, Moléculaire du Cycle Cellulaire, INSERM U309, Grenoble.
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 microL for a standard 3 x 160 x 0.5 mm strip), allowing micropreparative work (milligram samples) with a simple experimental design.
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