Method for rapid electrophoretic transfer of isoelectric focusing gels to polyvinylidene difluoride.
We present here a protocol for unidirectional wet electrotransfer of isoelectric focusing (IEF) gels to polyvinylidene difluoride (PVDF) that can be carried out in < 2 h, including preequilibration steps. In developing this protocol, we systematically evaluated sodium dodecyl sulfate and methanol concentrations in the preequilibration washes and length of electrotransfer. We demonstrated that the method is useful for tubulin, a protein that resists usual semi-dry transfer procedures due to its limited solubility under IEF conditions. We also successfully used our method for transferring the microtubule-associated protein, tau, as well as carbamylyte standards, creatine phosphokinase and carbonic anhydrase.
Lobert
S
S
School of Nursing, Department of Biochemistry, University of Mississippi Medical Center, Jackson 39216.
Correia
J J
JJ
D000180:Acrylic Resins; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005782:Gels; D007525:Isoelectric Focusing / Q000295:instrumentation*; D011145:Polyvinyls; D013997:Time Factors; D005465:Fluorocarbon Polymers
D000180:Acrylic Resins; D005782:Gels; D011145:Polyvinyls; C016680:polyacrylamide gels; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers