Macrophage migration inhibitory factor : methods and protocols [electronic resource]

  • Harris, James , Morand, Eric F.
  • Humana Press
  • 2020
Macrophage migration inhibitory factor : methods and protocols [electronic resource]


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 immunohishemistry; 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 prools, and tips on troubleshooting and avoiding known pitfalls.


1. Studying the Pro-Migratory Effects of MIF
Adrian Hoffmann, Leon Christian Zwißler, Omar El Bounkari, and Jürgen Bernhagen

2. In Vitro Approaches for Investigating the Influence of MIF on Leukocyte-Endothelial Cell Interactions under Flow Conditions
Michael J. Hickey

3. Using Intravital Microscopy to Study the Role of MIF in Leukocyte Trafficking In Vivo
M. Ursula Norman and Michael J. Hickey

4. Analysis of Secreted MIF from Cultured Murine Bone Marrow-Derived Immune Cells
Tali Lang

5. Measuring MIF in Biological Fluids
Fabien B. Vincent and Tali Lang

6. Flow Cytometry Phenotyping of Bone Marrow-Derived Macrophages from Wildtype and MIF KO Mice
Jacqueline K. Flynn, Nadia Deen, and James Harris

7. Genotyping Two Promoter Polymorphisms in the MIF Gene: a –794 CATT5-8 Microsatellite Repeat and a –173 G/C SNP
Lin Leng, Edwin Siu, and Richard Bucala

8. Staining MIF in Cells for Confocal Microscopy
James Harris

9. Microscopy Methods for Imaging MIF and its Interaction Partners
Kirstin D. Elgass, Sarah J. Creed, and Ina Rudloff

10. Co-Immunoprecipitation (Co-IP) of Macrophage Migration Inhibitory Factor (MIF)
Jawad H. Abidi, James Harris, and Nadia S. Deen

11. Concurrent Immunohishemical Localization and Western Blot Analysis of the MIF Receptor, CD74, in Formalin-Fixed, Paraffin-Embedded Tissue
Amanda Graham and Warren B. Nothnick

12. Methods to Determine the Effects of MIF on In Vitro Osteoclastogenesis using Murine Bone Marrow-Derived Cells and Human Peripheral Blood Mononuclear Cells (PBMCs)
Ran Gu

13. Inducing and Inhibiting Autophagy to Investigate its Interactions with MIF
Nadia S. Deen, Jacinta P. Lee, and James Harris

14. Assays for Measuring the Role of MIF in NLRP3 Inflammasome Activation
Anita Pinar and James Harris

15. Assays for Inducing and Measuring Cell Death to Detect Macrophage Migration Inhibitory Factor (MIF) Release
Shahrzad Zamani, Eric F. Morand, and Jacqueline K. Flynn

16. Assessing the Role of MIF in Plasmodium spp. Infections using Ex Vivo Models
Elizabeth H. Aitken

17. The Effect of Macrophage Migration Inhibitory Factor on Intestinal Permeability: FITC-Dextran Serum Measurement and Transmission Electron Microscopy
Milica Vujičić, Sanja Despotović, Tamara Saksida, and Ivana Stojanović

18. Investigating MIF in Mouse Models of Severe Corticosteroid-Resistant Neutrophilic Asthma
Venkata Sita Rama Raju Allam and Maria B. Sukkar

19. Investigating MIF in Mouse Model of GoutIzabela Galvão, Allysson Cramer, and Flavio Almeida Amaral

20. 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
Aiti Vizzini

21. Purification of Antibodies against Entamoeba histolytica MIF and their use in Analyzing Human and Mouse Samples Laura Farr, Koji Watanabe, and Shannon Moonah

22. Studying Plant MIF/D-DT-Like Genes and Proteins (MDLs)
Dzmitry Sinitski, Katrin Gruner, Jürgen Bernhagen, and Ralph Panstruga