Title: Adaptation of the equipment for high-performance electrophoresis to isoelectric focusing
Abstract: Fast and reproducible isoelectric focusing experiments have been performed in glass capillaries of 0.2 mm I.D., wall thickness 0.1 mm, and length 120 mm. These narrow, thin-walled tubes permit rapid removal of the Joule heat to allow high voltages (3000 V) and short run times. The experiments were carried out in equipment designed for high-performance electrophoresis. The protein zones were detected by on- or off-tube measurements of their UV absorption. This detection technique requires that the proteins can be mobilized following the isoelectric focusing step. This mobilization has been achieved either by pumping the zones past the stationary UV monitor (with the voltage applied to avoid zonal broadening during the elution) or by electrophoretic elution, accomplished by replacing the acid at the anode by a base (or the base at the cathode by an acid), which caused the pH gradient— and thereby the proteins— to move in the tube. The electrophoretic mobilization procedure is more universal because it is also applicable when the focusing is performed in a gel
Publication Year: 1985
Publication Date: 1985-01-01
Language: en
Type: article
Indexed In: ['crossref']
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Cited By Count: 427
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