Elution of glycoproteins from replicas of sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels.
A method for the elution of glycoproteins from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) replicas of gels on polyvinylidene difluoride (PVDF) membranes is described. Ten model glycoproteins were resolved by SDS-PAGE and then electrotransferred onto PVDF membranes. After reversible staining, glycoprotein bands were eluted with a mixture of SDS/Triton X-100 at pH 9 or with a mixture of guanidinium hydrochloride/lysophosphatidylcholine at neutral pH. For both types of eluents, the final recoveries ranged from over 30% to about 80%. Good recoveries and mild conditions of elution render the method applicable for the structural elucidation of glycan chains.
Szewczyk
B
B
Faculty of Biotechnology, University of Gdansk, Poland. szewczyk@biotech.univ.gda.pl
Pilat
Z
Z
Bienkowska-Szewczyk
K
K
Summers
D F
DF
D000180:Acrylic Resins; D000818:Animals; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D008567:Membranes, Artificial; D011145:Polyvinyls; D012967:Sodium Dodecyl Sulfate; D005465:Fluorocarbon Polymers
D000180:Acrylic Resins; D006023:Glycoproteins; D008567:Membranes, Artificial; D011145:Polyvinyls; D012967:Sodium Dodecyl Sulfate; C016680:polyacrylamide gels; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers