2017年8月15日
Top-down/Bottom-up Mass Spectrometry Workflow Using Dissolvable Polyacrylamide Gels.
Analytical chemistry
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- 巻
- 89
- 号
- 16
- 開始ページ
- 8244
- 終了ページ
- 8250
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1021/acs.analchem.7b00357
- 出版者・発行元
- AMER CHEMICAL SOC
Biologists' preeminent toolbox for separating, analyzing, and visualizing proteins is SDS-PAGE, yet recovering the proteins embedded in these polyacrylamide media as intact species is a long-standing challenge for mass spectrometry. In conventional workflows, protein mixtures from crude biological samples are electrophoretically separated at high-resolution within N,N'-methylene-bis-acrylamide cross-linked polyacrylamide gels to reduce sample complexity and facilitate sensitive characterization. However, low protein recoveries, especially for high molecular weight proteins, often hinder characterization by mass spectrometry. We describe a workflow for top-down/bottom-up mass spectrometric analyses of proteins in polyacrylamide slab gels using dissolvable, bis-acryloylcystamine-cross-linked polyacrylamide, enabling high-resolution protein separations while recovering intact proteins over a broad size range efficiently. The inferior electrophoretic resolution long associated with reducible gels has been overcome, as demonstrated by SDS-PAGE of crude tissue extracts. This workflow elutes intact proteins efficiently, supporting MS and MS/MS from proteins resolved on biologists' preferred separation platform.
- リンク情報
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- DOI
- https://doi.org/10.1021/acs.analchem.7b00357
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/28723075
- PubMed Central
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5590889
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000407988600011&DestApp=WOS_CPL
- ID情報
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- DOI : 10.1021/acs.analchem.7b00357
- ISSN : 0003-2700
- eISSN : 1520-6882
- PubMed ID : 28723075
- PubMed Central 記事ID : PMC5590889
- Web of Science ID : WOS:000407988600011