Related Experiment Videos
Isolation of aldehyde oxidase (EC 1.2.3.1) from potato extracts by preparative page
H L De Pooter1, Q L Huang, N M Schamp
1Laboratory of Organic Chemistry, Faculty of Agricultural Sciences, State University of Gent, Belgium.
Abstract:
A method is proposed for the isolation (and purification) of enzymes, with retention of their activity, from solutions or gels of preparative PAGE runs. It is based on the inclusion of Sephadex G-25 as a supporting medium for a collector buffer in otherwise normal disc-PAGE gels. The collector buffer has a lower pH and higher concentration than the stacking gel buffer. This makes the proteins concentrate in a very narrow, slowly moving band in the Sephadex on electrophoresis, and makes their recovery easy. The method is illustrated by the isolation of aldehyde oxidase from potato extracts (which was unsuccessful by classical methods), and of one isoenzyme from commercial lipoxygenase after preparative PAGE. Recovery of chicken egg albumin after PAGE was over 90%.