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Electrophoresis. 2010 Jan;31(3):440-7. doi: 10.1002/elps.200900435.

A multichannel gel electrophoresis and continuous fraction collection apparatus for high-throughput protein separation and characterization.

Author information

1
Genomics Division, Lawrence Berkeley National Laboratory, CA 94720, USA.

Abstract

To facilitate a direct interface between protein separation by PAGE and protein identification by mass spectrometry, we developed a multichannel system that continuously collects fractions as protein bands migrate off the bottom of gel electrophoresis columns. The device was constructed using several short linear gel columns, each of a different percent acrylamide, to achieve a separation power similar to that of a long gradient gel. A "Counter Free-Flow" elution technique then allows continuous and simultaneous fraction collection from multiple channels at low cost. We demonstrate that rapid, high-resolution separation of a complex protein mixture can be achieved on this system using SDS-PAGE. In a 2.5 h electrophoresis run, for example, each sample was separated and eluted into 48-96 fractions over a mass range of approximately 10-150 kDa; sample recovery rates were 50% or higher; each channel was loaded with up to 0.3 mg of protein in 0.4 mL; and a purified band was eluted in two to three fractions (200 microL/fraction). Similar results were obtained when running native gel electrophoresis, but protein aggregation limited the loading capacity to about 50 microg per channel and reduced resolution.

PMID:
20119951
DOI:
10.1002/elps.200900435
[Indexed for MEDLINE]

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