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.
CITATION STYLE
Greaser, M. L., & Warren, C. M. (2015). Method for resolution and western blotting of very large proteins using agarose electrophoresis. In Western Blotting: Methods and Protocols (pp. 285–291). Springer New York. https://doi.org/10.1007/978-1-4939-2694-7_30
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