Separation and purification of high molecular weight glycoproteins using agarose gel electrophoresis.
Several components of the extracellular matrix in the molecular weight range of 220 kDa to 150 kDa were purified by preparative electrophoresis on 2.5% Pro-Sieve agarose gels. These high molecular weight glycoproteins, separated under reducing conditions, were recovered in solution by extraction of individual agarose gel slices and analyzed on sodium dodecyl sulfate polyacrylamide gels and Western blots. This simple method permitted the separation and recovery of the laminin B chains (220 kDa and 205 kDa) and entactin (150 kDa) and may prove useful for the purification of other high molecular weight species.
Ugozzoli
M
M
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010.
Chiu
A
A
D000818:Animals; D015153:Blotting, Western; D004587:Electrophoresis, Agar Gel / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D016326:Extracellular Matrix Proteins / Q000302:isolation & purification*; D006023:Glycoproteins / Q000302:isolation & purification*; D007797:Laminin / Q000302:isolation & purification; D008562:Membrane Glycoproteins / Q000302:isolation & purification; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000302:isolation & purification; D009442:Neurilemmoma / Q000737:chemistry; D051381:Rats; D016622:Silver Staining; D014407:Tumor Cells, Cultured / Q000737:chemistry
D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D007797:Laminin; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C029967:nidogen