A Solubility Optimization Protocol for Two-Dimensional Gel Electrophoresis of Cardiac Tissue
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We outline a strategy for the optimization of buffer conditions for the solubilization, extraction, and isoelectric focusing
(IEF) of proteins from cardiac tissue for two-dimensional gel electrophoresis (2DE). This strategy, which involves altering
both the extraction and IEF buffers, allows one to ensure representation of the proteome that is as complete as possible.
Initial buffer choices are given, as well as basic protocols for modifications. Although these conditions have been effectively
demonstrated for human myocardium, in principle this procedure can be used for the initial screen of any new sample of tissue
or cultured cells.
Affiliation(s): (3) Department of Physiology, Queen’s University, Kingston, Ontario, Canada
(4) Department of Medicine, Johns Hopkins University, Baltimore, MD
(5) Department of Biological Chemistry, Johns Hopkins University, Baltimore, MD
(6) Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD
(4) Department of Medicine, Johns Hopkins University, Baltimore, MD
(5) Department of Biological Chemistry, Johns Hopkins University, Baltimore, MD
(6) Department of Biomedical Engineering, Johns Hopkins University, Baltimore, MD
Book Title: Cardiovascular Proteomics: Methods and Protocols
Series: Methods in Molecular Biology | Volume: 357 | Pub. Date: Feb-05-2007 | Page Range: 59-65 | DOI: 10.1385/1-59745-214-9:59
Subject: Molecular Medicine
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