The Protein Protocols Handbook / Edition 2

The Protein Protocols Handbook / Edition 2

by John M. Walker
ISBN-10:
0896039412
ISBN-13:
9780896039414
Pub. Date:
02/19/2002
Publisher:
Springer-Verlag New York, LLC

Paperback

View All Available Formats & Editions
Current price is , Original price is $99.0. You
Select a Purchase Option (2nd ed. 2002)
  • purchase options

Overview

The Protein Protocols Handbook / Edition 2

This greatly enhanced second edition introduces 60 critically important new chapters dealing with topics and techniques that have emerged since the first edition, as well as significantly updating the remaining articles. The new chapters cover many rapidly developing areas, particularly the application of mass spectrometry in protein characterization, as well as the now well-established 2-D PAGE technique in proteomics. The section on glycoprotein analysis has also been significantly expanded, and methods for the production of single-chain and phage-displayed antibodies have been added to the section on antibodies. Each readily reproducible method follows the highly praised format of the Methods in Molecular Biology™ series, offering a concise summary of its basic theory, a complete materials list, a step-by-step protocol for its successful execution, and extensive notes on avoiding pitfalls and on modifying the method to function within your own experimental circumstances.

Product Details

ISBN-13: 9780896039414
Publisher: Springer-Verlag New York, LLC
Publication date: 02/19/2002
Edition description: 2nd ed. 2002
Pages: 1176
Product dimensions: 7.01(w) x 10.00(h) x 0.07(d)

Table of Contents

Part I. Quantitation of Proteins
1 Protein Determination by UV Absorption
Alastair Aitken and Michèle P. Learmonth
2 The Lowry Method for Protein Quantitation
Jakob H. Waterborg
3 The Bicinchoninic Acid (BCA) Assay for Protein Quantitation
John M. Walker
4 The Bradford Method for Protein Quantitation
Nicholas J. Kruger
5 Ultrafast Protein Determinations Using Microwave Enhancement
Robert E. Akins and Rocky S. Tuan
6 The Nitric Acid Method for Protein Estimation in Biological Samples
Scott A. Boerner, Yean Kit Lee, Scott H. Kaufmann, and Keith C. Bible
7 Quantitation of Tryptophan in Proteins
Alastair Aitken and Michèle P. Learmonth
8 Flow Cytometric Quantitation of Cellular Proteins
Thomas D. Friedrich, F. Andrew Ray, Judith A. Laffin, and John M. Lehman
9 Kinetic Silver Staining of Proteins
Douglas D. Root and Kuan Wang
Part II. Electrophoresis of Proteins and Peptides and Detection in Gels
10 Nondenaturing Polyacrylamide Gel Electrophoresis of Proteins
John M. Walker
11 SDS Polyacrylamide Gel Electrophoresis of Proteins
John M. Walker
12 Gradient SDS Polyacrylamide Gel Electrophoresis of Proteins
John M. Walker
13 SDS-Polyacrylamide Gel Electrophoresis of Peptides
Ralph C. Judd
14 Identification of Nucleic Acid Binding Proteins Using Nondenaturing Sodium Decyl Sulfate Polyacrylamide Gel Electrophoresis (SDecS-Page)
Robert E. Akins and Rocky S. Tuan
15 Cetyltrimethylammonium Bromide Discontinuous Gel Electrophoresis of Proteins: Mr-Based Separation of Proteins with Retained Native Activity
Robert E. Akins and Rocky S. Tuan
16 Acetic–Acid–Urea Polyacrylamide Gel Electrophoresis of Basic Proteins
Jakob H. Waterborg
17 Acid–Urea–Triton Polyacrylamide Gel Electrophoresis of Histones
Jakob H. Waterborg
18 Isoelectric Focusing of Proteins in Ultra-Thin Polyacrylamide Gels
John M. Walker
19 Protein Solubility in Two-Dimensional Electrophoresis: Basic Principles and Issues
Thierry Rabilloud
20 Preparation of Protein Samples from Mouse and Human Tissues for 2-D Electrophoresis
Joachim Klose
21 Radiolabeling of Eukaryotic Cells and Subsequent Preparation for 2-D Electrophoresis
Nick Bizios
22 Two-Dimensional Polyacrylamide Gel Electrophoresis Using Carrier Ampholyte pH Gradients in the First Dimension
Patricia Gravel
23 Casting Immobilized pH Gradients (IPGs)
Elisabetta Gianazza
24 Nonequilibrium pH Gel Electrophoresis (NEPHGE)
Mary F. Lopez
25 Difference Gel Electrophoresis
Mustafa Ünlü and Jonathan Minden
26 Comparing 2-D Electrophoretic Gels Across Internet Databases
Peter F. Lemkin and Gregory C. Thornwall
27 Immunoblotting of 2-D Electrophoresis Separated Proteins
Barbara Magi, Luca Bini, Sabrina Liberatori, Roberto Raggiaschi, and Vitaliano Pallini
28 Quantification of Radiolabeled Proteins in Polyacrylamide Gels
Wayne R. Springer
29 Quantification of Proteins on Polyacrylamide Gels
Bryan John Smith
30 Rapid and Sensitive Staining of Unfixed Proteins in Polyacrylamide Gels with Nile Red
Joan-Ramon Daban, Salvador Bartolomé, Antonio Bermúdez, and F. Javier Alba
31 Zinc-Reverse Staining Technique
Carlos Fernandez-Patron
32 Protein Staining with Calconcarboxylic Acid in Polyacrylamide Gels
Jung-Kap Choi, Hee-Youn Hong, and Gyurng-Soo Yoo
33 Detection of Proteins in Polyacrylamide Gels by Silver Staining
Michael J. Dunn
34 Protein Detection on Polyacrylamide Gels and Electroblots Using Transition Metal Chelate Stains
Wayne F. Patton
35 Detection of Proteins in Polyacrylamide Gels by Fluorescent Staining
Michael J. Dunn
36 Detection of Proteins and Sialoglycoproteins in Polyacrylamide Gels Using Eosin Y Stain
Fan Lin and Gary E. Wise
37 Electroelution of Proteins from Polyacrylamide Gels
Paul Jenö and Martin Horst
38 Autoradiography and Fluorography of Acrylamide Gels
Antonella Circolo and Sunita Gulati
Part III. Blotting and Detection Methods
39 Protein Blotting by Electroblotting
Mark Page and Robin Thorpe
40 Protein Blotting by Semidry Method
Patricia Gravel
41 Protein Blotting by the Capillary Method
John M. Walker
42 Protein Blotting of Basic Proteins Resolved on Acid-Urea-Trinton-Polyacrylamide Gels
Geneviève P. Delcuve and James R. Davie
43 Alkaline Phosphatase Labeling of IgG Antibody
G. Brian Wisdom
44 b-Galactosidase Labeling of IgG Antibody
G. Brian Wisdom
45 Horseradish Peroxidase Labeling of IgG Antibody
G. Brian Wisdom
46 Digoxigenin (DIG) Labeling of IgG Antibody
G. Brian Wisdom
47 Conjugation of Fluorochromes to Antibodies
Su-Yau Mao
48 Coupling of Antibodies with Biotin
Rosaria P. Haugland and Wendy W. You
49 Preparation of Avidin Conjugates
Rosaria P. Haugland and Mahesh K. Bhalgat
50 MDPF Staining of Proteins on Western Blots
F. Javier Alba and Joan-Ramon Daban
51 Copper Iodide Staining of Proteins and Its Silver Enhancement
Douglas D. Root and Kuan Wang
52 Detection of Proteins on Blots Using Direct Blue 71
Hee-Youn Hong, Gyurng-Soo Yoo, and Jung-Kap Choi
53 Protein Staining and Immunodetection Using Immunogold
Susan J. Fowler
54 Detection of Polypeptides on Immunoblots Using Enzyme-Conjugated or Radiolabeled Secondary Ligands
Nicholas J. Kruger
55 Utilization of Avidin- or Streptavidin-Biotin as a Highly Sensitive Method to Stain Total Proteins on Membranes
Kenneth E. Santora, Stefanie A. Nelson, Kristi A. Lewis, and William J. LaRochelle
56 Detection of Protein on Western Blots Using Chemifluorescence
Catherine Copse and Susan J. Fowler
57 Quantification of Proteins on Western Blots using ECL
Joanne Dickinson and Susan J. Fowler
58 Reutilization of Western Blots After Chemiluminescent Detection or Autoradiography
Scott H. Kaufmann
Part IV. Chemical Modification of Proteins and Peptide Production and Purification
59 Carboxymethylation of Cysteine Using Iodoacetamide/Iodoacetic Acid
Alastair Aitken and Michèle P. Learmonth
60 Performic Acid Oxidation
Alastair Aitken and Michèle P. Learmonth
61 Succinylation of Proteins
Alastair Aitken and Michèle P. Learmonth
62 Pyridylethylation of Cysteine Residues
Malcolm Ward
63 Side Chain Selective Chemical Modifications of Proteins
Dan S. Tawfik
64 Nitration of Tyrosines
Dan S. Tawfik
65 Ethoxyformylation of Histidine
Dan S. Tawfik
66 Modification of Arginine Side Chains with p-Hydroxyphenylglyoxal
Dan S. Tawfik
67 Amidation of Carboxyl Groups
Dan S. Tawfik
68 Amidination of Lysine Side Chains
Dan S. Tawfik
69 Modification of Tryptophan with 2-Hydroxy-5-Nitrobenzylbromide
Dan S. Tawfik
70 Modification of Sulfhydryl Groups with DTNB
Dan S. Tawfik
71 Chemical Cleavage of Proteins at Methionyl-X Peptide Bonds
Bryan John Smith
72 Chemical Cleavage of Proteins at Tryptophanyl-X Peptide Bonds
Bryan John Smith
73 Chemical Cleavage of Proteins at Aspartyl-X Peptide Bonds
Bryan John Smith
74 Chemical Cleavage of Proteins at Cysteinyl-X Peptide Bonds
Bryan John Smith
75 Chemical Cleavage of Proteins at Asparaginyl-Glycyl Peptide Bonds
Bryan John Smith
76 Enzymatic Digestion of Proteins in Solution and in SDS Polyacrylamide Gels
Kathryn L. Stone and Kenneth R. Williams
77 Enzymatic Digestion of Membrane-Bound Proteins for Peptide Mapping and Internal Sequence Analysis
Joseph Fernandez and Sheenah Mische
78 Reverse Phase HPLC Separation of Enzymatic Digests of Proteins
Kathryn L. Stone and Kenneth R. Williams
Part V. Protein/Peptide Characterization
79 Peptide Mapping by Two-Dimensional Thin-Layer Electrophoresis–Thin-Layer Chromatography
Ralph C. Judd
80 Peptide Mapping by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis
Ralph C. Judd
81 Peptide Mapping by High-Performance Liquid Chromatography
Ralph C. Judd
82 Production of Protein Hydrolysates Using Enzymes
John M. Walker and Patricia J. Sweeney
83 Amino Acid Analysis by Precolumn Derivatization with 1-Fluoro-2,4-Dinitrophenyl-5-L-Alanine Amide (Marfey's Reagent)
Sunil Kochhar, Barbara Mouratou, and Philipp Christen
84 Molecular Weight Estimation for Native Proteins Using High-Performance Size-Exclusion Chromatography
G. Brent Irvine
85 Detection of Disulfide-Linked Peptides by HPLC
Alastair Aitken and Michèle P. Learmonth
86 Detection of Dis ulfide-Linked Peptides by Mass Spectrometry
Alastair Aitken and Michèle P. Learmonth
87 Diagonal Electrophoresis for Detecting Disulfide Bridges
Alastair Aitken and Michèle P. Learmonth
88 Estimation of Disulfide Bonds Using Ellman's Reagent
Alastair Aitken and Michèle P. Learmonth
89 Quantitation of Cysteine Residues and Disulfide Bonds by Electrophoresis
Alastair Aitken and Michèle P. Learmonth
90 Analyzing Protein Phosphorylation
John Colyer
91 Mass Spectrometric Analysis of Protein Phosphorylation
Débora Bonefant, Thierry Mini, and Paul Jenö
92 Identification of Proteins Modified by Protein (d-Aspartyl/l-Isoaspartyl) Carboxyl Methyltransferase
Darin J. Weber and Philip N. McFadden
93 Analysis of Protein Palmitoylation
Morag A. Grassie and Graeme Milligan
94 Incorporation of Radiolabeled Prenyl Alcohols and Their Analogs into Mammalian Cell Proteins: A Useful Tool for Studying Protein Prenylation
Alberto Corsini, Christopher C. Farnsworth, Paul McGeady, Michael H. Gelb, and John A. Glomset
95 The Metabolic Labeling and Analysis of Isoprenylated Proteins
Douglas A. Andres, Dean C. Crick, Brian S. Finlin, and Charles J. Waechter
96 2-D Phosphopeptide Mapping
Hikaru Nagahara, Robert R. Latek, Sergi A. Ezhevsky, and Steven F. Dowdy
97 Detection and Characterization of Protein Mutations by Mass Spectrometry
Yoshinao Wada
98 Peptide Sequencing by Nanoelectrospray Tandem Mass Spectrometry
Ole Nørregaard Jensen and Matthias Wilm
99 Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry for Protein Identification Using Peptide and Fragmention Masses
Paul L. Courchesne and Scott D. Patterson
100 Protein Ladder Sequencing
Rong Wang and Brian T. Chait
101 Sequence Analysis with WinGene/WinPep
Lars Hennig
102 Isolation of Proteins Cross-linked to DNA by Cisplatin
Virginia A. Spencer and James R. Davie
103 Isolation of Proteins Cross-linked to DNA by Formaldehyde
Virginia A. Spencer and James R. Davie
Part VI. Glycoproteins
104 Detection of Glycoproteins in Gels and Blots
Nicolle H. Packer, Malcolm S. Ball, Peter L. Devine, and Wayne F. Patton
105 Staining of Glycoproteins/Proteoglycans in SDS-Gels
Holger J. Møller and Jørgen H. Poulsen
106 Identification of Glycoproteins on Nitrocellulose Membranes Using Lectin Blotting
Patricia Gravel
107 A Lectin-Binding Assay for the Rapid Characterization of the Glycosylation of Purified Glycoproteins
Mohammad T. Goodarzi, Angeliki Fotinopoulou, and Graham A. Turner
108 Chemical Methods of Analysis of Glycoproteins
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
109 Monosaccharide Analysis by HPAEC
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
110 Monosaccharide Analysis by Gas Chromatography (GC)
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
111 Determination of Monosaccharide Linkage and Substitution Patterns by GC-MS Methylation Analysis
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
112 Sialic Acid Analysis by HPAEC-PAD
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
113 Chemical Release of O-Linked Oligosaccharide Chains
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
114 O-Linked Oligosaccharide Profiling by HPAEC-PAD
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
115 O-Linked Oligosaccharide Profiling by HPLC
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
116 Release of N-Linked Oligosaccharide Chains by Hydrazinolysis
Tsuguo Mizuochi and Elizabeth F. Hounsell
117 Enzymatic Release of O- and N-Linked Oligosaccharide Chains
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
118 N-Linked Oligosaccharide Profiling by HPLC on Porous Graphitized Carbon (PGC)
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
119 N-Linked Oligosaccharide Profiling by HPAEC-PAD
Elizabeth F. Hounsell, Michael J. Davies, and Kevin D. Smith
120 HPAEC-PAD Analysis of Monosaccharides Released by Exoglycosidase Digestion Using the CarboPac MA1 Column
Michael Weitzhandler Jeffrey Rohrer, James R. Thayer, and Nebojsa Avdalovic
121 Microassay Analyses of Protein Glycosylation
Nicky K. C. Wong, Nnennaya Kanu, Natasha Thandrayen, Geert Jan Rademaker, Christopher I. Baldwin, David V. Renouf, and Elizabeth F. Hounsell
122 Polyacrylamide Gel Electrophoresis of Fluorophore-Labeled Carbohydrates from Glycoproteins,
Brian K. Brandley, John C. Klock, and Christopher M. Star
123 HPLC Analysis of Fluorescently Labeled Glycans
Tony Merry
124 Glycoprofiling Purified Glycoproteins Using Surface Plasmon Resonance
Angeliki Fotinopoulou and Graham A. Turner
125 Sequencing Heparan Sulfate Saccharides
Jeremy E. Turnbull
126 Analysis of Glycoprotein Heterogeneity by Capillary Electrophoresis and Mass Spectrometry
Andrew D. Hooker and David C. James
127 Affinity Chromatography of Oligosaccharides and Glycopeptides with Immobilized Lectins
Kazuo Yamamoto, Tsutomu Tsji, and Toshiaki Osawa
Part. VII. Antibody Techniques
128 Antibody Production
Robert Burns
129 Production of Antibodies Using Proteins in Gel Bands
Sally Ann Amero, Tharappel C. James, and Sarah C. R. Elgin
130 Raising Highly Specific Polyclonal Antibodies Using Biocompatible Support-Bound Antigens
Monique Diano and André Le Bivic
131 Production of Antisera Using Peptide Conjugates
Thomas E. Adrian
132 The Chloramine T Method for Radiolabeling Protein
Graham S. Bailey
133 The Lactoperoxidase Method for Radiolabeling Protein
Graham S. Bailey
134 The Bolton and Hunter Method for Radiolabeling Protein
Graham S. Bailey
135 Preparation of I125 Labeled Peptides and Proteins with High Specific Activity Using IODO-GEN
J. Michael Conlon
136 Purification and Assessment of Quality of Radioiodinated Protein
Graham S. Bailey
137 Purification of IgG by Precipitation with Sodium Sulfate or Ammonium Sulfate
Mark Page and Robin Thorpe
138 Purification of IgG Using Caprylic Acid
Mark Page and Robin Thorpe
139 Purification of IgG Using DEAE-Sepharose Chromatography
Mark Page and Robin Thorpe
140 Purification of IgG Using Ion-Exchange HPLC
Carl Dolman, Mark Page, and Robin Thorpe
141 Purification of IgG by Precipitation with Polyethylene Glycol (PEG)
Mark Page and Robin Thorpe
142 Purification of IgG Using Protein A or Protein G
Mark Page and Robin Thorpe
143 Analysis and Purification of IgG Using Size-Exclusion High Performance Liquid Chromatography (SE-HPLC)
Carl Dolman and Robin Thorpe
144 Purification of IgG Using Affinity Chromatography on Antigen-Ligand Columns
Mark Page and Robin Thorpe
145 Purification of IgG Using Thiophilic Chromatography
Mark Page and Robin Thorpe
146 Analysis of IgG Fractions by Electrophoresis
Mark Page and Robin Thorpe
147 Purification of Immunoglobulin Y (IgY) from Chicken Eggs
Christopher R. Bird and Robin Thorpe
148 Affinity Purification of Immunoglobulins Using Protein A Mimetic (PAM)
Giorgio Fassina, Giovanna Palombo, Antonio Verdoliva, and Menotti Ruvo
149 Detection of Serological Cross Reactions by Western Cross Blotting
Peter Hammerl, Arnulf Hartl, Johannes Freund, and Josef Thalhammer
150 Bacterial Expression, Purification, and Characterization of Single-Chain Antibodies
Sergey M. Kipriyanov
151 Enzymatic Digestion of Monoclonal Antibodies
Sarah M. Andrew
152 How to Make Bispecific Antibodies
Ruth R. French
153 Phage Display: Biopanning on Purified Proteins and Proteins Expressed in Whole Cell Membranes
George K. Ehrlich, Wolfgang Berthold, and Pascal Bailon
154 Screening of Phage Displayed Antibody Libraries
Heinz Dörsam, Michael Braunagel, Christian Kleist, Daniel Moynet, and Martin Welschof
155 Antigen Measurements Using ELISA
William Jordan
156 Enhanced Chemiluminescence Immunoassay
Richard A. W. Stott
157 Immunoprecipitation
Kari Johansen and Lennart Svensson
Part VIII. Monoclonal Antibodies
158 Immunogen Preparation and Immunization Procedures for Rats and Mice,
Mark Page and Robin Thorpe
159 Hybridoma Production
Mark Page and Robin Thorpe
160 Screening Hybridoma Culture Supernatants Using Solid-Phase Radiobinding Assay
Mark Page and Robin Tho rpe
161 Screening Hybridoma Culture Supernatants Using ELISA
Mark Page and Robin Thorpe
162 Growth and Purification of Murine Monoclonal Antibodies
Mark Page and Robin Thorpe
163 Affinity Purification Techniques for Monoclonal Antibodies
Alexander Schwarz
164 A Rapid Method for Generating Large Numbers of High-Affinity Monoclonal Antibodies from a Single Mouse
Nguyen Thi Man and Glenn E. Morris

Customer Reviews

Most Helpful Customer Reviews

See All Customer Reviews