This second edition volume expands on the previous edition with new protocols that offer biochemical, biophysical, and biological assays for studying macrophage migration inhibitory factor (MIF) and D-dopachrome tautomerase (D-DT). The chapters in this book cover topics such as generation of MIF peptides via solid phase peptide synthesis; fragment screening of MIF by surface plasmon resonance; MIF NMR chemical shift perturbation mapping; methods to determine the role of MIF in cilia biogenesis; and immunofluorescent quantification of MIF in larval zebrafish tailfin transfection. 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. Cutting-edge and comprehensive, Macrophage Migration Inhibitory Factor: Methods and Protocols, Second Edition is a valuable resource for any researcher hoping to contribute to the ever expanding world of MIF biology.