Purification of rat C6 glioma plasma membranes by affinity partitioning.
We have studied the feasibility of purifying rat C6 glioma plasma membranes by a phase partitioning approach. The purification procedure involves cell homogenization and fractionation with an aqueous two-phase polymer system followed by selective affinity purification of plasma membranes by a wheat germ agglutinin-coupled polymer system. We demonstrate that the two-phase affinity partitioning technique is a simple and efficient method of isolating cell plasma membranes with high purity and yield. Furthermore, the isolated plasma membranes retain their functional integrity, as shown by the high-affinity insulin-like growth factor-I (IGF-I) binding capacity of IGF-I receptors.
Wang
Q
Q
Department of Biochemistry, University of Antwerp, Universitaire Instelling Antwerpen, Antwerp, Belgium. qwang@sickkids.on.ca
Wu
Y
Y
Aerts
T
T
Slegers
H
H
Clauwaert
J
J
D000345:Affinity Labels; D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D002498:Centrifugation; D003911:Dextrans; D005910:Glioma / Q000378:metabolism* / Q000648:ultrastructure*; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D011092:Polyethylene Glycols; D051381:Rats; D017526:Receptor, IGF Type 1 / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014909:Wheat Germ Agglutinins
D000345:Affinity Labels; D003911:Dextrans; D014909:Wheat Germ Agglutinins; D011092:Polyethylene Glycols; D007334:Insulin-Like Growth Factor I; D017526:Receptor, IGF Type 1