Electrotransfer of proteins in an environmentally friendly methanol-free transfer buffer.
Western blot transfer buffer was modified to substitute the acute poison methanol, with the common rubbing alcohol, isopropanol in concentrations of as low as 5 % for protein electrotransfer. Commercially available molecular weight markers and rabbit serum were run on polyacrylamide gels and shown to be transferred adequately to both nitrocellulose and polyvinylidene difluoride membranes under either wet or semi-dry conditions with similar results in all cases. This procedure was successfully used for immunodetection of the rabbit IgG heavy chain from serum. Therefore, this represents a good alternative for less toxic and environmentally friendly conditions for western immunoblotting of proteins.
Villanueva
Marco A
MA
Unidad Académica Puerto Morelos, Instituto de Ciencias del Mar y Limnología, Universidad Nacional Autónoma de México, Avenida Niños Héroes S/N, Puerto Morelos, Quintana Roo 77580, Mexico. marco@icmyl.unam.mx
D019840:2-Propanol; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002021:Buffers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007143:Immunoglobulin Heavy Chains / Q000302:isolation & purification*; D011817:Rabbits
D002021:Buffers; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D019840:2-Propanol