Rapid electrotransfer of proteins from polyacrylamide gel to nitrocellulose membrane using surface-conductive glass as anode.
A new type of inexpensive horizontal apparatus for the electrophoretic transfer of proteins from a gel to an immobilization membrane has been developed. In this system, gel and membrane were directly pressed between two flat electrodes. A surface-conductive glass was used as anode and a stainless-steel plate as cathode. Proteins could be transferred from polyacrylamide gel to nitrocellulose sheet, with a yield of at least 90% in 60-90 min, without overheating, using a voltage gradient of 30-40 V/cm. Moderate volumes of separate anodic (with 20% methanol) and cathodic (with 0.1% sodium dodecyl sulfate) buffers were suited for optimal transfer of proteins with relative molecular mass (Mr) in the 14,000-94,000 range.
D000818:Animals; D002021:Buffers; D002417:Cattle; D003101:Collodion; D004563:Electrochemistry; D004566:Electrodes; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation*; D005898:Glass; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000302:isolation & purification
D002021:Buffers; D011506:Proteins; D012710:Serum Albumin, Bovine; D003101:Collodion