A simplified method of protein electroblotting after isoelectric focusing on an ultrathin polyacrylamide gel layer fixed on a cellophane support.
A method is described for isoelectric focusing of proteins, using an ultrathin-layer polyacrylamide gel on cellophane, followed by electrophoretic transfer of separated proteins onto a nitrocellulose membrane. The polyacrylamide gel is firmly attached to the cellophane and thus protected from mechanical damage; such gels are easily manipulated. Cellophane is permeable to ions and application of this gel support overcomes difficulties resulting from the removal of ultrathin gels from a plastic support on electroblotting. Proteins separated under nondenaturing conditions were transferred onto a nitrocellulose membrane and detected by the concanavalin A-peroxidase technique. The proposed approach makes it possible to analyze the variability of nondenatured proteins and glycoproteins of different origin.
Yakhyayev
A V
AV
Research Center of Molecular Diagnostics, USSR Ministry of Health, Moscow.
Voronkova
I M
IM
Sukhanov
V A
VA
D000180:Acrylic Resins; D000818:Animals; D002476:Cellophane; D005782:Gels; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D008562:Membrane Glycoproteins / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D011506:Proteins / Q000032:analysis*; D051381:Rats; D011919:Rats, Inbred Strains
D000180:Acrylic Resins; D005782:Gels; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011506:Proteins; C016680:polyacrylamide gels; D002476:Cellophane