Subfractionation of Heart Tissue: The “In Sequence” Myofilament Protein Extraction of Myocardial Tissue
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Proteomic analysis of heart tissue is complicated by the large dynamic range of its proteins. The most abundant proteins are
the myofilament proteins, which comprise the contractile apparatus. This chapter describes a protocol for fractionation of
heart tissue that extracts the myofilament proteins into a separate sample fraction, allowing analysis of lower-abundance
proteins. Importantly, this is performed in a manner that is compatible with two-dimensional electrophoresis and high-performance
liquid chromatography, two of main technologies of proteomics. The method produces three fractions based on solubility at
different pHs: (1) cytoplasmic-enriched extract (neutral pH), (2) myofilament-enriched extract (acidic pH), and (3) membrane
protein-enriched pellet. Fractionation of heart tissue in this manner provides the basis for in-depth proteomic analysis.
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: 87-90 | DOI: 10.1385/1-59745-214-9:87
Subject: Molecular Medicine
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