Provides neutrophil protocols that are broadly applicable to leukocyte biology
Includes protocols for isolation of neutrophils from humans and animal species
Presents an interesting and informative overview of the field of neutrophil biology
Facilitates a better understanding of the role of neutrophils in host defense
This book provides a concise set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Scientists and clinicians will find this collection an invaluable aid.
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Neutrophil Methods and Protocols provides a concise set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Parts 1 & 2 provide an overview of neutrophils the methods most commonly used for isolating them from humans and other animal species. Part 3 details methods for investigating adhesion, chemotaxis, phagocytosis and bacterial activity. Part 4 presents protocols measuring gene expression in neutrophils, and Part 5 concludes with assays for diagnosing the most common neutrophil disorders. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.
Neutrophils Methods and Protocols Edited by Mark T. Quinn Frank R. DeLeo Gary M. Bokoch Contents Dedication i Preface ii Contributors v PART I: NEUTROPHILS: AN OVERVIEW 1. The Role of Neutrophils in the Immune System: An Overview Harry L. Malech PART II: NEUTROPHIL ISOLATION AND SUBCELLULAR FRACTIONATION 2. Isolation of Human Neutrophils From Venous Blood William M. Nauseef 3. Neutrophil Isolation From Non-Human Species Daniel W. Siemsen, Igor A. Schepetkin, Lilia N. Kirpotina, Benfang Lei, and Mark T. Quinn 4. Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers Lene Udby and Niels Borregaard PART III. BIOCHEMISTRY, BIOLOGY AND SIGNAL TRANSDUCTION OF NEUTROPHILS 5. Rac and Rap GTPase Activation Assays Ulla G. Knaus, Alison Bamberg, and Gary M. Bokoch 6. Measurement of Phospholipid Metabolism in Intact Neutrophils Susan Sergeant and Linda C. McPhail 7. Analysis of Protein Phosphorylation in Human Neutrophils Jamel El-Benna and Pham My-Chan Dang 8. Mitogen-Activated Protein Kinase Assays Martine Torres 9. Transduction of Proteins into Intact Neutrophils Tieming Zhao and Gary M. Bokoch 10. Optical Methods for the Measurement and Manipulation of Cytosolic Free Calcium in Neutrophils Esther J. Hillson, Sharon Dewitt, and Maurice B. Hallett 11. Analysis of Electrophysiological Properties and Responses of Neutrophils Deri Morgan and Thomas E. DeCoursey 12. Analysis of Neutrophil Apoptosis Emma L. Taylor, Adriano G. Rossi, Ian Dransfield, and Simon P. Hart PART IV. NEUTROPHIL ADHESION AND CHEMOTAXIS 13. Optical and Fluorescence Detection of Neutrophil Integrin Activation Ulrich Y. Schaff, Melissa R. Sarantos, Harold Ting, and Scott I. Simon 14. Analysis of Neutrophil Polarization and Chemotaxis Mary A. Lokuta, Paul A. Nuzzi, and Anna Huttenlocher 15. Quantifying and Localizing Actin Free Barbed Ends in Neutrophils Michael Glogauer 16. Measurement of Neutrophil Adhesion Under Conditions Mimicking Blood Flow Mark A. Jutila, Bruce Walcheck, Robert Bargatze, and Aiyappa Palecanda 17. Model Systems to Investigate Neutrophil Adhesion and Chemotaxis Nancy A. Louis, Eric Campbell, and Sean P. Colgan PART V. NEUTROPHIL PHAGOCYTOSIS AND BACTERICIDAL ACTIVITY 18. Immunofluorescence and Confocal Microscopy of Neutrophils Lee-Ann H. Allen 19. Assessment of Phagosome Formation and Maturation by Fluorescence Microscopy Benjamin E. Steinberg and Sergio Grinstein 20. Analysis of Neutrophil Membrane Traffic During Phagocytosis Per Lönnbro, Pontus Nordenfelt, and Hans Tapper 21. Analysis of Neutrophil Bactericidal Activity Jessie N. Green, Christine C. Winterbourn, and Mark B. Hampton 22. A Skin Chamber Technique as a Human Model for Studies of Aseptic Inflammatory Reactions Per Follin and Claes Dahlgren PART VI. NADPH OXIDASE AND PRODUCTION OF REACTIVE OXYGEN SPECIES 23. Measurement of Respiratory Burst Products Generated by Professional Phagocytes Claes Dahlgren, Anna Karlsson, and Johan Bylund 24. The K-562 Cell Model for Analysis of Neutrophil NADPH Oxidase Function Thomas L. Leto, Mark C. Lavigne, Neda Homoyounpour, Kristen Lekstrom, Gilda Linton, Harry L. Malech, and Isabelle de Mendez 25. Cell-free Assays: The Reductionist Approach to the Study of NADPH Oxidase Assembly Shahar Molshanski-Mor, Ariel Mizrahi, Yelena Ugolev, Iris Dahan, Yevgeny Berdichevsky, and Edgar Pick 26. Immunoaffinity Purification of Human Phagocyte Flavocytochrome b and Analysis
From the reviews:
"...a carefully compiled collection of protocols that allow for study of many aspects of the neutrophil." -Doody's Book Review, Weighted Numerical Score:85-3 Stars
"This excellent book is one of the latest in the `Methods in Molecular Biology´ series ... . it comes beautifully presented and bound in a hardback cover and would grace any bookshelf. I would urge all self-respecting laboratories working with neutrophils – whether in research or diagnostics – to purchase one immediately. ... I was impressed by this book´s comprehensive provision of protocols, addressing all aspects of neutrophil function and physiology, and including an excellent section on diagnosis of both inherited and non-inherited neutrophil disorders." (Lynn Morgan, Immunology News, May, 2008)
Neutrophil Methods and Protocols provides a concise set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Parts 1 & 2 provide an overview of neutrophils the methods most commonly used for isolating them from humans and other animal species. Part 3 details methods for investigating adhesion, chemotaxis, phagocytosis and bacterial activity. Part 4 presents protocols measuring gene expression in neutrophils, and Part 5 concludes with assays for diagnosing the most common neutrophil disorders. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.
Neutrophils: An Overview.- The Role of Neutrophils in the Immune System.- Neutrophil Isolation and Subcellular Fractionation.- Isolation of Human Neutrophils From Venous Blood.- Neutrophil Isolation From Nonhuman Species.- Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers.- Biochemistry, Biology, and Signal Transduction of Neutrophils.- Rac and Rap GTPase Activation Assays.- Measurement of Phospholipid Metabolism in Intact Neutrophils.- Analysis of Protein Phosphorylation in Human Neutrophils.- Mitogen-Activated Protein Kinase Assays.- Transduction of Proteins into Intact Neutrophils.- Optical Methods for the Measurement and Manipulation of Cytosolic Free Calcium in Neutrophils.- Analysis of Electrophysiological Properties and Responses of Neutrophils.- Analysis of Neutrophil Apoptosis.- Neutrophil Adhesion and Chemotaxis.- Optical and Fluorescence Detection of Neutrophil Integrin Activation.- Analysis of Neutrophil Polarization and Chemotaxis.- Quantifying and Localizing Actin-Free Barbed Ends in Neutrophils.- Measurement of Neutrophil Adhesion Under Conditions Mimicking Blood Flow.- Model Systems to Investigate Neutrophil Adhesion and Chemotaxis.- Neutrophil Phagocytosis and Bactericidal Activity.- Immunofluorescence and Confocal Microscopy of Neutrophils.- Assessment of Phagosome Formation and Maturation by Fluorescence Microscopy.- Analysis of Neutrophil Membrane Traffic During Phagocytosis.- Analysis of Neutrophil Bactericidal Activity.- A Skin Chamber Technique as a Human Model for Studies of Aseptic Inflammatory Reactions.- Nadph Oxidase and Production of Reactive Oxygen Species.- Measurement of Respiratory Burst Products Generated by Professional Phagocytes.- The K-562 Cell Model for Analysis of Neutrophil NADPH Oxidase Function.- Cell-Free Assays.- Immunoaffinity Purification of Human Phagocyte Flavocytochrome b and Analysis of Conformational Dynamics.- Analysis of Neutrophil Gene Expression and Tranillegalscription Factors.- Genome-Scale Tranillegalscript Analyses in Human Neutrophils.- Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Tranillegalscription Real-Time PCR.- Detection of Intact Tranillegalscription Factors in Human Neutrophils.- Neutrophil Defects and Diagnosis.- Disorders of Neutrophil Function.- Diagnostic Assays for Chronic Granulomatous Disease and Other Neutrophil Disorders.- Diagnostic Assays for Myeloperoxidase Deficiency.
From the reviews:
"...a carefully compiled collection of protocols that allow for study of many aspects of the neutrophil." -Doody's Book Review, Weighted Numerical Score:85-3 Stars
"This excellent book is one of the latest in the 'Methods in Molecular Biology' series ... . it comes beautifully presented and bound in a hardback cover and would grace any bookshelf. I would urge all self-respecting laboratories working with neutrophils - whether in research or diagnostics - to purchase one immediately. ... I was impressed by this book's comprehensive provision of protocols, addressing all aspects of neutrophil function and physiology, and including an excellent section on diagnosis of both inherited and non-inherited neutrophil disorders." (Lynn Morgan, Immunology News, May, 2008)
Inhaltsverzeichnis
Neutrophils
Methods and Protocols
Edited by
Mark T. Quinn
Frank R. DeLeo
Gary M. Bokoch
Contents
Dedication i
Preface ii
Contributors v
PART I: NEUTROPHILS: AN OVERVIEW
1. The Role of Neutrophils in the Immune System: An Overview
Harry L. Malech
PART II: NEUTROPHIL ISOLATION AND SUBCELLULAR FRACTIONATION
2. Isolation of Human Neutrophils From Venous Blood
William M. Nauseef
3. Neutrophil Isolation From Non-Human Species
Daniel W. Siemsen, Igor A. Schepetkin, Lilia N. Kirpotina, Benfang Lei, and Mark T. Quinn
4. Subcellular Fractionation of Human Neutrophils and Analysis of Subcellular Markers
Lene Udby and Niels Borregaard
PART III. BIOCHEMISTRY, BIOLOGY AND SIGNAL TRANSDUCTION OF NEUTROPHILS
5. Rac and Rap GTPase Activation Assays
Ulla G. Knaus, Alison Bamberg, and Gary M. Bokoch
6. Measurement of Phospholipid Metabolism in Intact Neutrophils
Susan Sergeant and Linda C. McPhail
7. Analysis of Protein Phosphorylation in Human Neutrophils
Jamel El-Benna and Pham My-Chan Dang
8. Mitogen-Activated Protein Kinase Assays
Martine Torres
9. Transduction of Proteins into Intact Neutrophils
Tieming Zhao and Gary M. Bokoch
10. Optical Methods for the Measurement and Manipulation of Cytosolic Free Calcium in Neutrophils
Esther J. Hillson, Sharon Dewitt, and Maurice B. Hallett
11. Analysis of Electrophysiological Properties and Responses of Neutrophils
Deri Morgan and Thomas E. DeCoursey
12. Analysis of Neutrophil Apoptosis
Emma L. Taylor, Adriano G. Rossi, Ian Dransfield, and Simon P. Hart
PART IV. NEUTROPHIL ADHESION AND CHEMOTAXIS
13. Optical and Fluorescence Detection of Neutrophil Integrin Activation
Ulrich Y. Schaff, Melissa R. Sarantos, Harold Ting, and Scott I. Simon
14. Analysis of Neutrophil Polarization and Chemotaxis
Mary A. Lokuta, Paul A. Nuzzi, and Anna Huttenlocher
15. Quantifying and Localizing Actin Free Barbed Ends in Neutrophils
Michael Glogauer
16. Measurement of Neutrophil Adhesion Under Conditions Mimicking Blood Flow
Mark A. Jutila, Bruce Walcheck, Robert Bargatze, and Aiyappa Palecanda
17. Model Systems to Investigate Neutrophil Adhesion and Chemotaxis
Nancy A. Louis, Eric Campbell, and Sean P. Colgan
PART V. NEUTROPHIL PHAGOCYTOSIS AND BACTERICIDAL ACTIVITY
18. Immunofluorescence and Confocal Microscopy of Neutrophils
Lee-Ann H. Allen
19. Assessment of Phagosome Formation and Maturation by Fluorescence Microscopy
Benjamin E. Steinberg and Sergio Grinstein
20. Analysis of Neutrophil Membrane Traffic During Phagocytosis
Per Lönnbro, Pontus Nordenfelt, and Hans Tapper
21. Analysis of Neutrophil Bactericidal Activity
Jessie N. Green, Christine C. Winterbourn, and Mark B. Hampton
22. A Skin Chamber Technique as a Human Model for Studies of Aseptic Inflammatory Reactions
Per Follin and Claes Dahlgren
PART VI. NADPH OXIDASE AND PRODUCTION OF REACTIVE OXYGEN SPECIES
23. Measurement of Respiratory Burst Products Generated by Professional Phagocytes
Claes Dahlgren, Anna Karlsson, and Johan Bylund
24. The K-562 Cell Model for Analysis of Neutrophil NADPH Oxidase Function
Thomas L. Leto, Mark C. Lavigne, Neda Homoyounpour, Kristen Lekstrom, Gilda Linton, Harry L. Malech, and Isabelle de Mendez
25. Cell-free Assays: The Reductionist Approach to the Study of NADPH Oxidase Assembly
Shahar Molshanski-Mor, Ariel Mizrahi, Yelena Ugolev, Iris Dahan, Yevgeny Berdichevsky, and Edgar Pick
26. Immunoaffinity Purification of Human Phagocyte Flavocytochrome b and Analysis
Klappentext
Neutrophil Methods and Protocols provides a concise set of protocols for assessing basic neutrophil functions, investigating specialized areas in neutrophil research, and completing step-by-step diagnostic assays of common neutrophil disorders. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. Parts 1 & 2 provide an overview of neutrophils the methods most commonly used for isolating them from humans and other animal species. Part 3 details methods for investigating adhesion, chemotaxis, phagocytosis and bacterial activity. Part 4 presents protocols measuring gene expression in neutrophils, and Part 5 concludes with assays for diagnosing the most common neutrophil disorders. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.
Provides neutrophil protocols that are broadly applicable to leukocyte biology
Includes protocols for isolation of neutrophils from humans and animal species
Presents an interesting and informative overview of the field of neutrophil biology
Facilitates a better understanding of the role of neutrophils in host defense
