Abstract
Very large proteins (subunit sizes, >200 kDa) are difficult to electrophoretically separate on polyacrylamide gels. A SDS vertical agarose gel system has been developed that has vastly improved resolving power for very large proteins. Proteins with molecular masses between 200 and 4000 kDa can be clearly separated. Inclusion of a reducing agent in the upper reservoir buffer and use of a large pore-sized agarose have been found to be key technical procedures for obtaining optimum protein migration and resolution.
Key words
- SeaKem Gold agarose
- Titin
- DATD
- Large proteins
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Acknowledgments
This work was supported by the College of Agricultural and Life Sciences, University of Wisconsin-Madison, and from grants MLG- NIH HL77196 and Hatch NC1131.
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Greaser, M.L., Warren, C.M. (2019). Electrophoretic Separation of Very Large Molecular Weight Proteins in SDS Agarose. In: Kurien, B., Scofield, R. (eds) Electrophoretic Separation of Proteins. Methods in Molecular Biology, vol 1855. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8793-1_18
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DOI: https://doi.org/10.1007/978-1-4939-8793-1_18
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