Cardiovascular Proteomics: Methods and Protocols presents cutting-edge protocols and strategies for proteomic evaluation of cardiovascular disease written by pioneering researchers in the field. Topics explored in this comprehensive volume include obtaining specific heart proteins, cutting-edge techniques for identifying risk biomarkers of atherome plaque rupture, analyzing the secretome of explanted endarterectomies cultured in vitro, and phage display techniques for deciphering the molecular diversity of blood vessels. Detailed protocols for the isolation of short- and long-term culture of the adult mouse cardiac myocytes are also included. Numerous advanced proteomic techniques are addressed, including protein separation by two-dimensional electrophoresis and two-dimensional difference gel electrophoresis, liquid chromatography coupled to mass spectrometry, and the SELDI-TOF approach to searching for biomarkers of stroke in human serum or of hypertension in the serum of animal models. Proteomic strategies and protocols for studying the proteome of endothelial, arterial smooth muscle cells, foam cells, and circulating blood monocytes constitute a major element of this text. Cutting-edge techniques for the analysis of subproteomes from isolation to final characterization and the characterization of posttranslational modifications are also addressed. This useful snapshot of current proteomic techniques is a single source for protocols for the identification and validation of novel biomarkers and targets in cardiovascular diseases.
...a fine collection of methods and protocols currently employed in cardiovascular proteomics that will be of great use to all who are invovled in cardiovascular research. -Proteomics Clin. Appl. "...a thorough set of protocols for the cardiac biology community. Most laboratories will have some of these in place, but can profit from the broad coverage of this book." -Doody's Book Review, Weighted Numerical Score: 76 - 3 stars
Part I. Analysis of Heart Proteome Two-dimensional Polyacrylamide Gel Electrophoresis for Cardiovascular Proteomics Michael J. Dunn Analyzing The Cardiac Muscle Proteome By Liquid Chromatography-Mass Spectrometry (Lc-ms) Based Expression Proteomics Anthony O. Gramolini, Thomas Kislinger, Peter Liu, David H. Maclennan, and Andrew Emili 2-D Differential Gel Electrophoresis (DIGE) of Rat Heart Proteins in Ischemia and Ischemia-Reperfusion Jun Sakai, Hironori Ishikawa, Hideshi Satoh, Setsuko Yamamoto, Shinichi Kojima, and Masaharu Kanoaka Analysis of Antihypertensive Drugs in the Heart of Animal Models: A Proteomic Approach Alberto Lazaro, Julio Gallego-delgado, Julio Osende, Jesus Egido, and Fernando Vivanco A Solubility Optimization Protocol for 2-D Gel Electrophoresis of Cardiac Tissue Brian A. Stanley and Jennifer E. Van Eyk A Method for Effective Depletion of Albumin From Cellular Extracts: Application to Human Myocardium Brian A. Stanley and Jennifer E. Van Eyk Part II. Heart Subproteomes Subcellular Fractionation Mariana E. Guimaraes De Araujo and Alfons Huber Subfractionation of Heart Tissue: The "In Sequence" Myofilament Protein Extraction Of Myocardial Tissue Lesley A. Kane, Irina Neverova, and Jennifer E. Van Eyk Optimization of Cardiac Troponin I Pull-Down by Idm Affinity Beads and Seldi-tofT Diane E. Bovenkamp, Brian A. Stanley and Jennifer E.van Eyk Proteomic Analysis Of The Subunit Composition of Complex I (Nadh:Ubiquinone Oxidoreductase) From Bovine Heart Mitochondria Ian M. Fearnley, Joe Carrol and John E. Walker Identification of Targets of Phosphorylation in Heart Mitochondria Roberta A. Gottliebie Part III. Proteomics of Human Atheroma Plaques Characterization of The Human Atheroma Plaque Secretome by Proteomic Analysis Mari-Carmen Duran, Jose L. Martin-Ventura, Sebastian Mas, Maria G. Barderas, Veronica M. Darde, Ole N. Jensen, Jesus Egido, and Fernando Vivanco Characterization of Hsp-27 Phosphorylation Sites in Human Atherosclerotic Plaque Secretome Mari-Carmen Duran, Elisabetta Boeri-Erba, Shabaz Mohammed, Jose L. Martin-Ventura, Jesus Egido, Fernando Vivanco, and Ole N. Jensen Western Array Analysis of Human Atherosclerotic Plaque Wim Martinetar Part IV. The Proteome of Cells of The Cardiovascular System The Proteome of Endothelial Cells Jesus Gonzalez-Cabrero, Mayte Pozo, Mari-Carmen Duran, Rosario de Nicolas, Jesus Egido, and Fernando Vivanco Proteomic Study of Caveolae and Rafts Isolated from Human Endothelial Cells Richard R. Sprenger and Anton J. G. Horrevoets Proteomic Identification of S-nitrosylated Proteins In Endothelial Cells Antonio Martinez-Ruiz and Santiago Lamasgo The Proteome and Secretome of Human Arterial Smooth Muscle Cell Annabelle Dupont and Florence Pinet Real-Time in Vivo Proteomic Identification Of Novel Kinase Substrates in Smooth Muscle Anne A. Wooldridge and Timothy A. Haystead Proteomic Analysis of Vascular Smooth Muscle Cells Treated With Ouabain Alexy V. Pshezhetsky Isolation and Culture of Adult Mouse Cardiac Myocytes Timothy D. O'Connell, Manoj C. Rodrigo, and Paul C. Simpson Proteomic Analysis Of Foam Cells Peng-Yuang Yang, Yao-cheng Rui, Peng-Yuan Yang, and Yan-Ling Yuin Isolation of The Platelet Releasate Judith Coppinger, Desmond J. Fitzgerald, and Patricia B. Maguire Enrichment Of Phospho-Tyrosine Proteome Of Human Platelets By Immunoprecipitation Martina Foy, Donald F. Harney, Kieran Wynne, and Patricia B. Maguire Chapter 25 Veronica M. Darde, Mari-Carmen Duran, Jesus Egido, and Fernando Vivanco Part V. Cardiovascular Biomarkers in Plasma Cardiovascular Biomarkers Discovery By Seldi-tof Mass Spectrometry Olivier Meilhac, Sandrine Delbosc, and Jean-Baptiste Michel Detection of Biomarkers of Stroke Using Seldi-tofe Jean-charles Sanchez, Pierre Lescuyer, Denis Hochstrasser, and Laure Allard Depletion of High-Abundance Proteins In Plasma by Immunoaffinity Substraction For Two-Dimensional Difference Gel Electrophoresis (DIGE) Analysis Veronica M. Darde, Maria G. Barderas, and Fernando Vivanco A Rapid, Economic and Reproducible Method For Human Serum Delipidation, Albumin And IgG Removal, For Proteomic Analysis Qin Fu, Diane E. Bovenkamp And Jennifer E.van Eyk Part VI. Bioinformatics: Emerging Technologies Mitoproteome: Human Heart Mitochondrial Protein Sequence Database Purnima Guda, Shankar Subramaniam, and Chittibabu Guda Techniques To Decipher Molecular Diversity By Phage Display Dawn R. Christianson, Michael G. Ozawa, Renata Pasqualini, and Wadih Arapraprap
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