Full Description
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.
Contents
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 Transcription Factors.- Genome-Scale Transcript Analyses in Human Neutrophils.- Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Transcription Real-Time PCR.- Detection of Intact Transcription 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.
