Isoelectric focusing of B-lactamases on cellulose acetate membranes

Abstract
Cellulose acetate membranes treated with boron trifluoride in methanol were found to provide suitable support material for isoelectric focusing of β-lactamases; the method used is described. A linear pH gradient was obtained and the application of visible pi markers allowed the pH gradient to be monitored as focusing proceeded. β-Lactamases extracted from antibiotic-resistant coliform organisms focused as sharp, straight bands upon visualization with nitrocefin. Results were known within two hours of applying the β-lactamase extracts. The method allows a very economical use of expensive ampholyte and offers a quick and more simple alternative procedure for the identification of β-lactamases than the conventional polyacrylamide gel method.

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