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Updated: Apr 11, 2026

Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins
Published on: June 14, 2016
Method for resolution and western blotting of very large proteins using agarose electrophoresis
Marion L Greaser1, Chad M Warren
1Muscle Biology Laboratory, University of Wisconsin-Madison, 1805 Linden Drive, Madison, WI, 53706, USA, mgreaser@ansci.wisc.edu.
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.
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