内容

目次

Part I: Quantitation of Proteins. Protein Determination by UVAbsorption, Alastair Aitken and Michele P. Learmonth. The Lowry Method forProtein Quantitation, Jakob H. Waterborg. The Bicinchoninic Acid (BCA) Assayfor Protein Quantitation, John M. Walker. The Bradford Method for ProteinQuantitation, Nick J. Kruger. Ultrafast Protein Determinations UsingMicrowave Enhancement, Robert E. Akins and Rocky S. Tuan. The Nitric AcidMethods for Protein Estimation in Biological Samples, S.A. Boerner, Y.K. Lee,S.H. Kaufmann, and Keith C. Bible. Quantitation of Tryptophan in Proteins,Alastair A, Aitken and Michele P. Learmonth. Flow Cytometric Quantitation ofCellular Proteins, Thomas D. Friedrich, F. Andrew Ray, Judith A. Laffin, andJohn M. Lehman. Kinetic Silver Staining of Proteins, Douglas D. Root and KuanWang. Part II: Electrophoresis of Proteins and Peptides and Detection inGels. Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins, John M.Walker. SDS Polyacrylamide Gel Electrophoresis of Proteins, John M. Walker.Gradient SDS Polyacrylamide Gel Electrophoresis of Proteins, John M. Walker.SDS-Polyacrylamide Gel Electrophoresis of Peptides, Ralph C. Judd.Identification of Nucleic Acid Binding Proteins Using Nondenaturing SDS-PAGE,Robert E. Aitkens and Rocky S. Tuan. Cetyltrimethylammonium BromideDiscontinuous Gel Electrophoresis of Proteins: M-Based Separation of Proteinswith Retained Native Activity, Robert E. Aitkens and Rocky S. Tuan.Acetic-Acid-Urea Polyacrylamide Gel Electrophoresis of Basic Proteins, JakobH. Waterborg. Acid-Urea-Triton Polyacrylamide Gel Electrophoresis ofHistones, Jakob H. Waterborg. Isoelectric Focusing of Proteins in Ultra-ThinPolyacrylamide Gels, John M. Walker. Protein Solubility in 2DElectrophoresis: Basic Principles and Issues, Thierry Rabilloud. Preparationof Protein Samples from Mouse and Human Tissues for 2-D Electrophoresis,Joachim Klose. Radiolabelling of Eukaryotic Cells and Subsequent Preparationfor 2-D, Nick Bizios. Two-Dimensional PAGE Using Carrier Ampholyte pHGradients in the First Dimension, Patricia Gravel. Casting Immobilized pHGradients, Elisabetta Gianazza. Nonequilibrium pH Gel Electrophoresis, MaryLopez. Difference Gel Electrophoresis, Mustafa Unlu. Comparing 2-DElectrophoresis Gels Across Internet Databases, Peter F. Lemkin.Immunoblotting of 2-DE Separated Proteins, Barbara Magi, Luca Bini, SabrinaLiberatori, Roberto Raggiarchi, and Vitaliano Pallini. Quantification ofRadiolabeled Proteins in Polyacrylamide Gels, Wayne R. Springer.Quantification of Proteins on Polyacrylamide Gels, Bryan J. Smith. Rapid andSensitive Staining of Unfixed Proteins in Polyacrylamide Gels with Nile Red,John-Ramon Daban. Zinc Reverse Staining Technique, Carlos Fernandez-Patron.Protein Staining with Calconcarboxylic Acid in Polyacrylamide Gels, Jung-KapChoi. Detection of Proteins in Polyacrylamide Gels by Silver Staining,Michael J. Dunn. Protein Detection on Polyacrylamide Gels and on ElectroblotsUsing Transition Metal Chelate Stains, Wayne F. Patton. Detection of Proteinsin Polyacrylamide Gels by Fluorescent Staining, Michael J. Dunn. Detection ofProteins and Sialogycoproteins in Polyacrylamide Gels Using Eosin X Stain,Fan Lin and Gary Wise. Electroelution of Proteins from Polyacrylamide Gels,Paul Jeno. Autoradiography and Fluorography of Acrylamide Gels, AntonellaCircolo. Part III: Blotting and Detection of Proteins. Protein Blotting byElectroblotting, Robin Thorpe. Protein Blotting by the Semidry Method,Patricia Gravel. Protein Blotting by the Capillary Method, John M. Walker.Protein Blotting of Basic Proteins Resolved onAcid-Urea-Trinton-Polyacrylamide Gels, Genevieve P. Delcuve and James R.Davie. Alkaline Phosphatase Labeling of IgG Antibody, G. Brian Wisdom.b-Galactosidase Labeling of IgG Antibody, G. Brian Wisdom . HorseradishPeroxidase Labeling of IgG Antibody, G. Brian Wisdom. Digoxigenin (DIG)Labeling of IgG Antibody, G. Brian Wisdom. Conjugation of Fluorochromes toAntibodies, Su-Yau Mao. Coupling of Antibodies with Biotin, Rosaria P.Haugland. Preparation of Avidin Conjugates, Rosaria P. Haugland and M.K.Bhalgat. MDPF Staining of Proteins on Western Blots, F. Alba and Joan-RamonDaban. Cooper Iodided Staining of Proteins and its Silver Enhancement,Douglas D. Root and Kuan Wang. Detection of Proteins on Blots Using DirectBlue 71, Hee-Youn Hong, Gyurng-Soo Yoo, and Jung-Kap Choi. Protein Stainingand Immunodetection Using Immunogold, Susan J. Fowler. Detection ofPolypeptides on Immunoblots Using Enzyme Conjugated or Radiolabeled SecondaryLigands, Nicholas J. Kruger. Avidin- or Streptavidin-Biotin as a HighlySensitive Method to Stain Total Proteins on Membranes, Kenneth E. Santora,Stefanie A. Nielson, Kristi A. Lewis, and William J. LaRochelle. Detection ofProtein on Blots Using Chemifluorescence, Catherine C. Copse and Susan J.Fowler. Quantification of Proteins on Western Blots using ECL, J. Dickinsonand Susan J. Fowler. Reutilization of Western Blots After ChemiluminescentDetection or Autoradiography, Scott Kaufmann. Part IV: Chemical Modificationof Proteins and Peptide Production and Purification. Carboxymethylation ofCysteine Using Iodoacetamide/Iodoacetic Acid, Alastair Aitken. Performic AcidOxidation, Alastair Aitken. Succinylation of Protein, Alastair Aitken.Pyridylethylation of Cystenine Residues, Malcolm Ward. Side-Chain SelectiveChemical Modifications of Proteins, Dan S. Tawfik. Nitration of Tyrosines,Dan S. Tawfik. Ethoxyformylation of Histidine, Dan S. Tawfik. Modification ofArginine Side Chains with p-Hydroxyphenylglyoxal, Dan S. Tawfik. Amidinationof Carboxyl Groups, Dan S. Tawfik. Amidination of Lysine Side Chains, Dan S.Tawfik. Modification of Tryptophan with 2-Hydroxy-5 Nitrogenzylbromide, DanS. Tawfik. Modification of Sulhydryl Groups with DTNB, Dan S. Tawfik.Chemical Cleavage of Proteins at Methionyl-X Peptide Bonds., Bryan J. Smith.Chemical Cleavage of Proteins at Trypotophanyl-X Peptide Bonds, Bryan J.Smith. Chemical Cleavage of Proteins at Aspartyl-X Peptide Bonds, Bryan J.Smith. Chemical Cleavage of Proteins at Cysteinyl-X Peptide Bonds, Bryan J.Smith. Chemical Cleavage of Proteins at Asparaginyl-Glycyl Peptide Bonds,Bryan J. Smith. Enzymatic Digestion of Proteins in Solution and in SDSPolyacrylamide Gels , Kathryn L. Stone. Enzymatic Digestion of Proteins onPVDF Membranes, Joseph Fernandez and Sheenah Mische. Reverse Phase HPLCSeparation of Enzymatic Digests of Proteins, Kathryn L. Stone. Part V:Protein/Peptide Characterization. Peptide Mapping by two-DimensionalThin-Layer Electrophoresis-Thin-Layer Chromatography, Ralph C. Judd. PeptideMapping by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, RalphC. Judd. Peptide Mapping by High-Performance Liquid Chromatography, Ralph C.Judd. Production of Protein Hydrolysates Using Enzymes, John M. Walker andPatricia J. Sweeney. Amino Acid Analysis by Pre-Column Derivatization,Philipp Christen, Molecular Weight Estimation for Native Proteins UsingHigh-Performance Size Exclusion Chromatography, G. Brent Irvine. Detection ofDisulfide-Linked Peptides by HPLC, Alastair Aitken and Michele P. Learmonth.Detection of Disulfide-Linked Peptides by Mass Spectrometry, Alastair Aitkenand Michele P. Learmonth. Diagonal Electrophoresis for Detection ofDisulfide-Bridges, Alastair Aitken and Michele P. Learmonth. Estimation ofDisulfide Bonds Using Ellman's Reagent, Alastair Aitken and Michele P.Learmonth. Quantitation of Cysteine Residues and Disulfide Bonds byElectrophoresis, Alastair Aitken and Michele P. Learmonth. Analyzing ProteinPhosphorylation John Colyer x 91 Mass Spectrometric Analysis of ProteinPhosphorylation Debora Bonefant, Thierry Mini, and Paul Jeno x 92Identification of Proteins Modified by Protein Carboxyl MethyltransferaseDarin J. Weber and Philip N. McFadden x 93 Analysis of Protein PalmitoylationMorag A. Grassie and Graeme Milligan x 94 Incorporation of RadiolabeledPrenyl Alcohols and Their Analogs into Mammalian Cell Proteins: A Useful Toolfor Studying Protein Prenylation Alberto Corsini x 95 The MetabolicLabelling and Analysis of Isoprenylated Proteins Douglas Andres, Dean Crick,Brian Finlin, and Charles Waechter x 96 Two-Dimensional Phosphopeptidemapping Steven F. Dowdy x 97 Detection and Characterization of ProteinMutations by Mass Spectrometry Yoshinao Wada x 98 Peptide sequencing byNanoelectrospray Tendem Mass Spectrometry Ole Norregaard Jensen x 99 MALDIMass Spectrometry for Protein Identification Using Peptide and Fragment IonMasses Paul Courchesne and Scott D. Patterson x 100 Protein Ladder SequencingBrian T. Chait x 101 Sequence Analysis with WinGene/WinPep Lars Hennig x 101Sequence Analysis with WinGene/WinPep Lars Hennig x 102 Isolation of ProteinsCross-Linked to DNA by Cisplatin Virginia A. Spencer and James R. Davie x 103Isolation of Proteins Cross-Linked to DNA by Formaldehyde Virginia A. Spencerand James R. Davie x Part VI : Glycoproteins 104 Detection of Glycoproteinsin Gels and Blots Nicolle H. Packer, Malcolm Ball, Peter L. Devine, andWayne, F. Patton x 105 Staining of Glycoproteins/Proteoglycans on SDS-GelsHolger J. Moller x 106 Identification of Glycoproteins on NitrocelluloseMembranes Using Lectin Blotting Patricia Gravel x 107 A Lectin-Binding Assayfor the Rapid Characterization of the Glycosylation of Purified GlycoproteinsMohammad T. Goodarzi, Angeliki Fontinopoulou, and Graham A. Turner x 108Chemical Methods of Analysis of Glycoproteins Elizabeth F. Hounsell, MichaelJ. Davies, and Kevin D. Smith x 109 Monosaccharide Analysis by HPAECElizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith x 110Monosaccharide Analysis by GC Elizabeth F. Hounsell, Michael J. Davies, andKevin D. Smith x 111 Determination of Monosaccharide Linkage and SubstitutionPatterns by GC,MS Methylation Analysis Elizabeth F. Hounsell, Michael J.Davies, and Kevin D. Smith x 112 Sialic Acid Analysis by HPAEC-PAD ElizabethF. Hounsell, Michael J. Davies, and Kevin D. Smith x 113 Chemical Release ofO-Linked Oligosaccharide Chains Elizabeth F. Hounsell, Michael J. Davies, andKevin D. Smith x 114 O-Linked Oligosaccharide Profiling by HPLC Elizabeth F.Hounsell, Michael J. Davies, and Kevin D. Smith x 115 O-LinkedOligosaccharide Profiling by HPLC Elizabeth F. Hounsell, Michael J. Davies,and Kevin D. Smith x 116 Release of N-Linked Oligosaccharide Chains byHydrazinolysis T. Mizuochoi and Elizabeth F. Hounsell x 117 EnzymaticRelease of O- and N-Linked Oligosaccharide Chains Elizabeth F. Hounsell,Michael J. Davies, and Kevin D. Smith x 118 N-Linked OligosaccharideProfiling by HPLC on Porous Graphitized Carbon (PGC) Elizabeth F. Hounsell,Michael J. Davies, and Kevin D. Smith x 119 N-Linked OligosaccharideProfiling by HPLC on Porous Graphitized Carbon (PGC) Elizabeth F. Hounsell,Michael J. Davies, and Kevin D. Smith x 120 N-Linked OligosaccharideProfiling by HPAEC-PAD Michael Weitzhandler x 121 Microassay Analysis ofProtein Glycosylation Nicky K.C. Wong x 122 Polyacrylamide GelElectrophoresis of Fluorophore-Labeled Carbohydrates from Glycoproteins BrianK. Brandley, John C. Klock, and Christopher M. Star x 123 HPLC Analysis ofFluorescently Labelled Glycans Tony Merry x 124 Glycoprofiling PurifiedGlycoproteins using Surface Plasmon Resonance Angeliki Fotinopoulou and G.A.Turner x Sequencing Heparan Sulphate Saccharides Jeremy E. Turnbull x 126Analysis of Glycoprotein Heterogeneity by CE and Mass Spectrometry Andrew D.Hooker and David C. James x 127 Affinity Chromatography of Oligosaccharidesand Glycopeptides with Immobilized Lectins Kazuo Yamamoto x Part VII :Antibody Production 128 Antibody Production Robert Burns x 129 Production ofAntibodies Using Proteins in Gel Bands Sally Ann Amero, Tharappel C. James,and Sarah C.R. Elgin x 130 Raising Highly Specific Polyclonal AntibodiesUsing Biocompatible Support Bound Antigens Monique Diano and Andre Le Bivicx 131 Production of Antisera Using Peptide Conjugates Thomas E. Adrian x132 The Chloramine T Method for Radiolabeling Protein Graham S. Bailey x 133The Lactoperoxidase Method for Radiolabeling Protein Graham S. Bailey x 134The Bolton and Hunter Method for Radiolabeling Protein Graham S. Bailey x135 Preparation of I125 Labelled Peptides and Proteins With High SpecificActivity Using Iodogen J. Michael Conlon x 136 Purification and Assessment ofQuality of Radioiodinated Protein Graham S. Bailey x 137 Purification of IgGby Precipitation with Sodium Sulfate or Ammonium Sulfate Mark Page and RobinThorpe x 138 Purification of IgG Using Caprylic Acid Mark Page and RobinThorpe x 139 Purification of IgG Using DEAE-Sepharose Chromatography MarkPage and Robin Thorpe x 140 Purification of IgG Using Ion-Exchange HPLC MarkPage and Robin Thorpe 141 Purification of IgG by Precipitation withPolyethylene Glycol (PEG) Mark Page and Robin Thorpe x 142 Purification ofIgG Using protein A or Protein G Mark Page and Robin Thorpe x 143Purification of IgG Using Size Exclusion High Performance LiquidChromatography Carl Dolman Robin Thorpe x 144 Purification of IgG UsingAffinity Chromatography on Antigen-Ligand Columns Mark Page and Robin Thorpex 145 Purification of IgG Using Thiophilic Chromatography Mark Page andRobin Thorpe x 146 Analysis of IgG Fractions by Electrophoresis Mark Pageand Robin Thorpe x 147 Purification of Immunoglobulin Y (IgY) from ChickenEggs C.R. Bird and Robin Thorpe x 148 Affinity Purification ofImmunoglobulins Using PAM (Protein A Mimetic) Giorgio Fassina x 149Detection of Serological Cross Reactions by Western Cross Blotting JosefThalhammer x 150 Bacterial Expression, Purification, and Characterization ofSingle-Chain Antibodies Sergey M. Kipriyanovx 151 Enzymatic Digestion ofMonoclonal Antibodies Sarah M. Andrew x 152 How to make Bispecific AntibodiesRuth R. French x 153 Phage Display: Biopanning on Purified Proteins andProteins Expressed in Whole Cell Membranes George Ehrlich, Wolfgang Berthold,and Pascal Bailon x 154 Screening of Phage Displayed Antibody LibrariesMartin Welschof x 155 Antigen Measurements Using ELISA William Jordan x 156Enhanced Chemiluminescence Immunoassay Richard A.W. Scott x 157Immunoprecipitation Kari Johansen x Part VIII: Monoclonal antibodies 158Immunogen Preparation and Immunization Procedures for Rats and Mice Mark Pageand Robin Thorpe x 159 Hybridoma Production Mark Page and Robin Thorpe x 160Screening Hybridoma Culture Supernatants Using Solid-Phase RadiobindingAssay Mark Page and Robin Thorpe x 161 Screening Hybridoma CultureSupernatants Using ELISA Mark Page and Robin Thorpe x 162 Growth andPurification of Murine Monoclonal Antibodies Mark Page and Robin Thorpe x 163Affinity Purification Techniques for Monoclonal Antibodies Mark Page andRobin Thorpe x 164 A Rapid Method for Generating Large Numbers ofHigh-Affinity Monoclonal Antibodies from a Single Mouse N. Man and Glenn E.Morris x Index