Journal
ELECTROPHORESIS
Volume 33, Issue 2, Pages 316-324Publisher
WILEY-BLACKWELL
DOI: 10.1002/elps.201100364
Keywords
Membrane proteome; Protein fractionation; Sample cleanup; SDS; Special gradient gel
Funding
- Hunan Provincial Natural Science Foundation of China [11JJ2019]
- Specialized Research Foundation for the doctorial Program of Higher Education of China [20094306110003]
- National Natural Science Foundation of China [31070700]
- National Basic Research Program or 973 Program of China [2010CB529800, 2007CB914203]
Ask authors/readers for more resources
SDS is mostly used to enhance the solubilization and extraction of membrane proteins due to its strong detergency and low cost. Nevertheless, SDS interferes with the subsequent procedures and needs to be removed from the samples. In this work, a special gradient gel electrophoresis (GGE) system was developed to remove SDS from the SDS-solubilized protein samples. As a proof-of-principle experiment, the GGE system was designed to be composed of an agarose loading layer, six polyacrylamide fractionation layers with different concentrations and a high-concentration polyacrylamide sealing layer. The advantages of the GGE system are that it not only can electrophoretically remove SDS efficiently so that the protein loss resulted from the repeated gel washing after electrophoresis was avoided, but also can reduce the complexity of the sample, prevent the precipitation of proteins after loading and avoid the loss of proteins with low molecular weight during the electrophoresis. Using GGE system, about 85% of SDS in the sample and gel was electrophoretically removed and the proteins were fractionated. Compared with the two representative gel-based sample cleanup methods reported in literature, GGE-based strategy significantly improved the identification efficiency of proteins in terms of the number and coverage of the identified proteins.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available