Guide to protein purification / edited by Murray P. Deutscher.

Includes bibliographical references and indexes.

Print version record.

Guide to Protein Purification, designed to serve the needs of the student, experienced researcher and newcomer to the field, is a comprehensive manual that provides all the up-to-date procedures necessary for purifying, characterizing, and handling proteins and enzymes in one source. Key Features * Detailed procedures newly written for this volume * Extensive practical information * Rationale and strategies for protein and enzyme purification * Personal perspectives on enzyme purification by eminent researchers Among the Topics Covered * General methods for handling proteins and enzymes * Extraction, subcellular fractionation, and solubilization procedures * Comprehensive purification techniques * Specialized purification procedures * Protein characterization * Immunological procedures * Computer analysis of protein structure.

Why purify enzymes / Arthur Kornberg -- Strategies and considerations for protein purifications / Stuart Linn -- Setting up a laboratory / Murray P. Deutscher -- Buffers: Principles and practice / Vincent S. Stoll, John S. Blanchard -- Measurement of enzyme activity / Edward F. Rossomando -- Quantitation of protein / Christa M. Stoscheck -- Concentration of proteins and removal of solutes / Thomas Pohl -- Maintaining protein stability / Murray P. Deutscher -- Overproduction of proteins in Escherichia coli: Vectors, hosts, and strategies / Asis Das -- Overexpression of proteins in eukaryotes / Margaret K. Bradley -- Secretion of heterologous proteins in Escherichia coli / I. Barry Holland, Brendan Kenny, Boris Steipe, Andreas Plückthun -- Preparation of extracts from prokaryotes / Millard Cull, Charles S. McHenry -- Preparation of extracts from yeast / S. Michal Jazwinski -- Preparation of Extracts from Plants / Peter Gegenheimer -- Preparation of extracts from higher eukaryotes / John David Dignam -- Isolation of subcellular organelles / Brian Storrie, Edward A Madden -- Preparation of membrane fractions / Juris Ozols -- Detergents: An overview / Judith M. Neugebauer -- Solubilization of native membrane proteins / Leonard M. Hjelmeland -- Solubilization of protein aggregates / Fiona A.O. Marston, Donna L. Hartley -- Removal of detergents from membrane proteins / Leonard M. Hjelmeland -- Precipitation techniques / Sasha Englard, Sam Seifter -- Precipitation of proteins with polyethylene glycol / Kenneth C. Ingham -- Ion-exchange chromatography / Edward F. Rossomando -- Gel filtration / Earle Stellwagen -- Protein chromatography on hydroxyapatite columns / Marina J. Gorbunoff -- Hydrophobic chromatography / Robert M. Kennedy -- Chromatography on immobilized reactive dyes / Earle Stellwagen -- Affinity chromatography: General methods / Steven Ostrove -- Affinity chromatography: Specialized techniques / Steven Ostrove, Shelly Weiss -- Chromatofocusing / Lallan Giri -- High-performance liquid chromatography: Effective protein purification by various chromatographic modes / Roman M. Chicz, Fred E. Regnier -- One-dimensional gel electrophoresis / David E. Garfin -- Protein analysis using high-resolution two-dimensional polyacrylamide gel electrophoresis / Bonnie S. Dunbar, Hitomi Kimura, Therese M. Timmons -- Isoelectric focusing / David E. Garfin -- Gel-staining techniques / Carl R. Merril -- Elution of protein from gels / Michael G. Harrington -- Purification of membrane proteins / Thomas C. Thomas, Mark G. McNamee -- Purification of DNA-binding proteins by site-specific DNA affinity chromatography / Shlomo Eisenberg, Stephen C. Francesconi, Chris Civalier, Scott S. Walker -- Purification of glycoproteins / Craig Gerard -- Purification of multienzyme complexes / Paul A. Srere, Christopher K. Mathews -- Determination of purity / David G. Rhodes, Thomas M. Laue -- Determination of size, molecular weight, and presence of subunits / Thomas M. Laue, David G. Rhodes -- Amino acid analysis / Juris Ozols -- Limited N-terminal sequence analysis / Paul Matsudaira -- Peptide mapping / Ralph C. Judd -- Analysis for protein modifications and nonprotein cofactors / Sam Seifter, Sasha Englard -- Protein crystallization / David Ollis, Stephen White -- Preparation of polyclonal antibodies / Bonnie S. Dunbar, Eric D. Schwoebel -- Preparation of monoclonal antibodies / Bonnie S. Dunbar, Sheri M. Skinner -- Protein blotting and immunodetection / Therese M. Timmons, Bonnie S. Dunbar -- Immunoprecipitation of proteins / Gary L. Firestone, Sandra D. Winguth -- Immunoassays / Charles W. Parker -- Radiolabeling of proteins / Charles W. Parker -- Using purified protein to clone its gene / John M. Wozney -- Computer analysis of protein structure / Patrick Argos -- Rethinking your purification procedure / Murray P. Deutscher -- The private life of DNA polymerase I / Arthur Kornberg -- Purification and crystallization of transaldolase from Candida utilis / B.L. Horecker, O. Tsolas -- Discovery of glutamine synthetase cascade / Earl R. Stadtman -- Discovery, resolution, purification, and function of elongation factors / Kivie Moldave.

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