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This second edition provides updated and new chapters to build on and extend the strengths of the first edition. Chapters guide readers through basic biology of basophils, obtaining the cells by purification, culture of stem cells progenitors, peripheral CD34+ stem cell-derived mast cells, basophils from CD34+ progenitors, diagnostic applications, gene expression patterns in basophils, roles of basophils in different asthma phenotypes, knockout, and disease models. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Basophils and Mast Cells: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.
List of contents
Paradigm Shifts in Mast Cell and Basophil Biology and Function: An Emerging View of Immune Regulation in Health and Disease.- Purification of Basophils from Peripheral Human Blood.- Mast Cell Purification Protocols.- Generation of a Human Allergic Mast Cell Phenotype from CD133+ Stem Cells.- Generation and Culture of Peripheral CD34+ Stem Cell-derived Mast Cells (PSCMCs).- Modulating the Human Basophil Phenotype During its Development and Maturation: Basophils Derived from in vitro Cultures of CD34+ Progenitor Cells.- Generation of Mast Cells from Murine Stem Cell Progenitors.- Integration of the Human Dermal Mast Cell into the Organotypic Co-culture Skin Model.- The Absolute Basophil Count.- Mast Cell and Basophils Cell Lines: A Compendiu.- Use of Humanized RBL Reporter Systems for Detection of Allergen-specific IgE Sensitization in Human Serum.- Use of Humanized Fluorescent Reporter Cell Line RBL NFAT-DsRed for Detection of Allergen-specific IgE in Patient Sera Using Allergen Microarrays.- NPY-mRFP Rat Basophil Leukemia (RBL) Reporter: A Novel, Fast Reporter of Basophil/Mast Cell Degranulation.- Use of Engineered Nanoparticles (ENPs) for the Study of High-affinity IgE Receptor Fc RI Receptor Engagement and Rat Basophil Leukemia (RBL) Cell degranulation.- Flow Cytometric Allergy Diagnosis: Basophil Activation Techniques.- Basophil Activation Experiments in Immediate Drug Hypersensitivity: More Than a Diagnostic Aid.- Basophil Activation Techniques: Staining of Exteriorized Granule Matrix.- Measurement and Functional Analysis of the Mas-related G protein-coupled receptor MRGPRX2 on Human Mast Cells and Basophils.- The Mast Cell Activation Test (MAT).- Quantity and Quality of Basophil RNA Depend on the RNA Extraction Technique.- Measuring Histamine and Cytokine Release from Basophils and Mast Cells.- Detection of Mast Cells and Basophils by Immunohistochemistry.- Detection of Basophils and Other Granulocytes in Induced Sputum by Flow Cytometry.- Mast Cell Migration and Chemotaxis Assayed by Microscopy.- Basophil Stimulation and Signaling Pathway.- Gene Silencing Approaches in Mast Cells and Primary Human Basophils.- Identification and Immunophenotypic Characterization of Normal and Pathological Mast Cells.- Mast Cell Mediated Reactions in vivo.- Identification of Murine Basophils by Flow Cytometry and Histology.
