A multi-channel gel electrophoresis and continuous fraction collection apparatus for high throughput protein separation and characterization

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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 ... continued below

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Choi, Megan; Nordmeyer, Robert A.; Cornell, Earl; Dong, Ming; Biggin, Mark D. & Jin, Jian October 2, 2009.

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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 10-150 kDa; sample recovery rates were 50percent 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 L/fraction). Similar results were obtained when running native gel electrophoresis, but protein aggregation limited the loading capacity to about 50 g per channel and reduced resolution.

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  • Journal Name: Electrophoresis; Journal Volume: 31; Journal Issue: 3; Related Information: Journal Publication Date: January 2010

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  • Report No.: LBNL-3280E
  • Grant Number: DE-AC02-05CH11231
  • Office of Scientific & Technical Information Report Number: 983162
  • Archival Resource Key: ark:/67531/metadc1012178

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Office of Scientific & Technical Information Technical Reports

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  • October 2, 2009

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  • Oct. 14, 2017, 8:36 a.m.

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  • Oct. 17, 2017, 6:22 p.m.

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Choi, Megan; Nordmeyer, Robert A.; Cornell, Earl; Dong, Ming; Biggin, Mark D. & Jin, Jian. A multi-channel gel electrophoresis and continuous fraction collection apparatus for high throughput protein separation and characterization, article, October 2, 2009; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc1012178/: accessed October 21, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.