Preparative acrylamide electrophoresis: A single gel system

Abstract

1. 1. An apparatus is described for continuous elution of substances as they are separated by electrophoresis in acrylamide gel. The elution buffer used for this purpose is pumped inward across the base of a cylinder of gel and then passes upward and out via a central capillary. 2. 2. Proteins and other large molecules are prevented from passing into the electrode buffer by means of a cellophane membrane. The membrane is rigidly mounted on a Perspex grid which forms part of an elution cell. 3. 3. Bubble formation in the membrane support grid is prevented by a magnetic stirrer operating in the lower electrode chamber. 4. 4. With 0.05 M borate buffer at pH 9.2 in the electrode vessels and for elution, and 0.37 M Tris buffer at pH 8.8 in the gel, substances with mobilities down to one-half of that of albumin can be separated and eluted from the gel during 24 hr of electrophoresis.