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Concurrent Immunohistochemical Localization and Western Blot Analysis of the MIF Receptor, CD74, in Formalin-Fixed, Paraffin-Embedded Tissue

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Macrophage Migration Inhibitory Factor

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2080))

Abstract

Archived, formalin-fixed, paraffin-embedded tissue provides a robust resource for assessing protein expression in a variety of complex tissue types. Immunohistochemical localization techniques allow one to identify proteins of interest in the different cell populations which compose these tissues, but quantitative comparison within and between samples is semiquantitative. In contrast, Western blot analysis provides a more quantitative assessment but without the ability to identify the cellular sources of expressed protein. Here we describe a dual approach using human endometrium to assess both the localization and quantitation of the macrophage migration inhibitory factor (MIF) receptor CD74 by immunohistochemical techniques and Western blotting.

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References

  1. David JR (1966) Delayed hypersensitivity in vitro: its mediation by cell-free substances formed by lymphoid cell-antigen interaction. Proc Natl Acad Sci U S A 56:72–77

    Article  CAS  Google Scholar 

  2. Bloom BR, Bennett B (1966) Mechanism of a reaction in vitro associated with delayed-type hypersensitivity. Science 153:80–88

    Article  CAS  Google Scholar 

  3. Leng L, Metz CN, Fang Y, Xu J, Donnelly S, Baugh J, Delohery T, Chen Y, Mitchell RA, Bucala R (2003) MIF signal transduction initiated by binding to CD74. J Exp Med 197:1467–1476

    Article  CAS  Google Scholar 

  4. Bozzi F, Mogavero A, Varinelli L, Belfiore A, Manenti G, Caccia C, Volpi CC, Beznoussenko GV, Milione M, Leoni V, Gloghini A, Mironov AA, Leo E, Pilotti S, Pierotti MA, Bongarzone I, Gariboldi M (2017) MIF/CD74 axis is a target for novel therapies in colon carcinomatosis. J Exp Clin Cancer Res 36:16

    Article  Google Scholar 

  5. McClelland M, Zhao L, Carskadon S, Arenberg D (2009) Expression of CD74, the receptor for macrophage migration inhibitory factor, in non-small cell lung cancer. Am J Pathol 174:638–646

    Article  CAS  Google Scholar 

  6. Takahashi K, Koga K, Linge HM, Zhang Y, Lin X, Metz CN, Al-Abed Y, Ojamaa K, Miller EJ (2009) Macrophage CD74 contributes to MIF-induced pulmonary inflammation. Respir Res 10:33

    Article  Google Scholar 

  7. Valiño-Rivas L, Baeza-Bermejillo C, Gonzalez-Lafuente L, Sanz AB, Ortiz A, Sanchez-Niño MD (2015) CD74 in kidney disease. Front Immunol 26:48

    Google Scholar 

  8. Nothnick WB, Falcone T, Olson MR, Fazleabas AT, Tawfik OW, Graham A (2018) Macrophage migration inhibitory factor receptor, CD74 is overexpressed in human and baboon (Papio Anubis) endometriotic lesions and modulates endometriotic epithelial cell survival and interleukin-8 expression. Reprod Sci 11:1557–1166

    Article  Google Scholar 

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Correspondence to Warren B. Nothnick .

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Graham, A., Nothnick, W.B. (2020). Concurrent Immunohistochemical Localization and Western Blot Analysis of the MIF Receptor, CD74, in Formalin-Fixed, Paraffin-Embedded Tissue. In: Harris, J., Morand, E. (eds) Macrophage Migration Inhibitory Factor. Methods in Molecular Biology, vol 2080. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9936-1_11

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  • DOI: https://doi.org/10.1007/978-1-4939-9936-1_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9935-4

  • Online ISBN: 978-1-4939-9936-1

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