Full Description
This volume looks at--and discusses the techniques of--a range of areas in biology in which macrophage migration inhibitory factor (MIF) is studied. Topics covered in this book include methods to quantify MIF in cell supernatants and human samples, using ELISA and western blot; detection and analysis of intracellular MIF using flow cytometry; genotyping of human MIF polymorphisms, detection of MIF expression in a marine invertebrate using rapid amplification of cDNA RACE; studying the MIF receptor CD74 in tissues using immunohistochemistry; and studying MIF in models of disease such as asthma, Plasmodium infection, and gout. 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.
Comprehensive and practical, Macrophage Migration Inhibitory Factor: Methods and Protocols is a valuable resource that will help researchers gain a new understanding of MIF biology in health and disease.
Contents
Studying the Pro-Migratory Effects of MIF.- In Vitro Approaches for Investigating the Influence of MIF on Leukocyte-Endothelial Cell Interactions under Flow Conditions.- Using Intravital Microscopy to Study the Role of MIF in Leukocyte Trafficking In Vivo.- Analysis of Secreted MIF from Cultured Murine Bone Marrow-Derived Immune Cells.- Measuring MIF in Biological Fluids.- Flow Cytometry Phenotyping of Bone Marrow-Derived Macrophages from Wildtype and MIF KO Mice.- Genotyping Two Promoter Polymorphisms in the MIF Gene: a -794 CATT5-8 Microsatellite Repeat and a -173 G/C SNP.- Staining MIF in Cells for Confocal Microscopy.- Microscopy Methods for Imaging MIF and its Interaction Partners.- Co-Immunoprecipitation (Co-IP) of Macrophage Migration Inhibitory Factor (MIF).- Concurrent Immunohistochemical Localization and Western Blot Analysis of the MIF Receptor, CD74, in Formalin-Fixed, Paraffin-Embedded Tissue.- Methods to Determine the Effects of MIF on In Vitro Osteoclastogenesis using Murine Bone Marrow-Derived Cells and Human Peripheral Blood Mononuclear Cells (PBMCs).- Inducing and Inhibiting Autophagy to Investigate its Interactions with MIF.- Assays for Measuring the Role of MIF in NLRP3 Inflammasome Activation.- Assays for Inducing and Measuring Cell Death to Detect Macrophage Migration Inhibitory Factor (MIF) Release.- Assessing the Role of MIF in Plasmodium spp. Infections using Ex Vivo Models.- The Effect of Macrophage Migration Inhibitory Factor on Intestinal Permeability: FITC-Dextran Serum Measurement and Transmission Electron Microscopy.- Investigating MIF in Mouse Models of Severe Corticosteroid-Resistant Neutrophilic Asthma.- Investigating MIF in Mouse Model of Gout.- 5'and 3' RACE Method to Obtain Full-Length 5' and 3' Ends of Ciona robusta Macrophage Migration Inhibitory Factors MIF 1 and MIF 2 cDNA.- Purification of Antibodies against Entamoeba histolytica MIF and their use in Analyzing Human and Mouse Samples.- Studying Plant MIF/D-DT-Like Genes and Proteins (MDLs).
