title: Method for resolution and western blotting of very large proteins using agarose electrophoresis.
abstract: Proteins larger than 200 kDa are difficult to separate electrophoretically using polyacrylamide gels, and their transfer during western blotting is typically incomplete. A vertical SDS agarose gel system was developed that has vastly improved resolving power for very large proteins. Complete transfer of proteins as large as titin (Mr 3,000-3,700 kDa) onto blots can be achieved. The addition of a sulfhydryl reducing agent in the upper reservoir buffer and transfer buffer markedly improves the blotting of large proteins.
authors.affiliation: Muscle Biology Laboratory, University of Wisconsin-Madison, 1805 Linden Drive, Madison, WI, 53706, USA, mgreaser@ansci.wisc.edu.
authors.forename: Marion L
authors.initials: ML
authors.lastname: Greaser
authors.forename: Chad M
authors.initials: CM
authors.lastname: Warren
mesh_terms: D015153:Blotting, Western / Q000379:methods*; D002021:Buffers; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D013194:Staining and Labeling
chemical_list: D002021:Buffers; D011506:Proteins