title: Specific detection of interferon regulatory factor 5 (IRF5): A case of antibody inequality.
abstract: Interferon regulatory factor 5 (IRF5) is a member of the IRF family of transcription factors. IRF5 was first identified and characterized as a transcriptional regulator of type I interferon expression after virus infection. In addition to its critical role(s) in the regulation and development of host immunity, subsequent studies revealed important roles for IRF5 in autoimmunity, cancer, obesity, pain, cardiovascular disease, and metabolism. Based on these important disease-related findings, a large number of commercial antibodies have become available to study the expression and function of IRF5. Here we validate a number of these antibodies for the detection of IRF5 by immunoblot, flow cytometry, and immunofluorescence or immunohistochemistry using well-established positive and negative controls. Somewhat surprising, the majority of commercial antibodies tested were unable to specifically recognize human or mouse IRF5. We present data on antibodies that do specifically recognize human or mouse IRF5 in a particular application. These findings reiterate the importance of proper controls and molecular weight standards for the analysis of protein expression. Given that dysregulated IRF5 expression has been implicated in the pathogenesis of numerous diseases, including autoimmune and cancer, results indicate that caution should be used in the evaluation and interpretation of IRF5 expression analysis.
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.|Rutgers Biomedical and Health Sciences, New Jersey Medical School-Cancer Center, Newark, NJ 07103, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.|Rutgers Biomedical and Health Sciences, New Jersey Medical School-Cancer Center, Newark, NJ 07103, United States.|Department of Microbiology, Biochemistry &Molecular Genetics, Rutgers Biomedical and Health Sciences, Newark, NJ 07103, United States.
authors.forename: Saurav
authors.initials: S
authors.lastname: De
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.
authors.forename: Su
authors.initials: S
authors.lastname: Song
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.
authors.forename: Bharati
authors.initials: B
authors.lastname: Matta
authors.affiliation: Center for Autoimmune and Musculoskeletal Diseases, Feinstein Institute for Medical Research, North well Health, Manhasset, NY 11030, United States.|Rutgers Biomedical and Health Sciences, New Jersey Medical School-Cancer Center, Newark, NJ 07103, United States.
authors.forename: Betsy J
authors.initials: BJ
authors.lastname: Barnes
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D002478:Cells, Cultured; D050835:Interferon Regulatory Factors / Q000276:immunology / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009000:Monocytes / Q000276:immunology* / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D050835:Interferon Regulatory Factors; C498178:Irf5 protein, mouse