Fast electrotransfer of human serum proteins in their native state from polyacrylamide thin gradient gels reinforced by textiles to polyvinylidene difluoride membranes. Rapid electrotransfer from thin gradient gels reinforced by textiles.
A fast electroblotting technique of native molecules electrophoretically separated in thin (0.25 to 0.5 mm) gradient gels, onto a high capacity membrane of polyvinylidene difluoride is described. Omitting methanol during transfer, the equilibration step is avoided and the same buffer is used in electrophoresis and transfer. As the gel reinforced by fabric never swells nor shrinks, and as all the bands are blotted, the transfer matrix exactly reflects the protein pattern of the original gel. Autoradiography is enhanced and electroelution is homogeneous in all parts of the gels. Significant improvement is noticed in binding proteins of molecular weight from about 20 kDa to more than 700 kDa, as suggested by complete electroelution of all native serum components.
Laboratoire de Biochimie, Faculté de Médecine, Université de Nice-Sophia Antipolis, France.
M
M
Starita-Geribaldi
P
P
Sudaka
D001798:Blood Proteins / Q000302:isolation & purification*; D004563:Electrochemistry; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D008567:Membranes, Artificial; D011145:Polyvinyls; D005465:Fluorocarbon Polymers
D001798:Blood Proteins; D008567:Membranes, Artificial; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers