AGAR ELECTROPHORESIS Part V. Preparative Agar Electrophoresis of Serum Proteins

Abstract

Agar electrophoresis developed by Giri has been adapted for preparative purposes. The method consists in preparing agarbuffer gel in a perspex through in the centre of which a slit is made for the protein solution under test. About 1 to 2 C.C, of human serum has been subjected to preparative agar electrophoresis and separated into four distinct fractions aIbumin ((2, f3 and y-globulin, which havebeen isolated in a homogeneous state. The protein is eluted from the agar gel by freezing at-20°C for 3 hours and thawing it at room temperature. The resulting solution is filtered free of agar on a sintered glass funnel, dialysed free of buffersalts and then concentrated under vacuo over phosphorus pentoxide in cold. The isolated fractions retain their original mobility without getting denatured. The method can be used for the isolation of protein components in the native state ona semi- reparative scale. The recovery of protein from the agar gel is of the order of 95 per cent as shown by recovery experiments done with pure albumin.