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

Methods Mol Biol. 2020:2080:123-134. doi: 10.1007/978-1-4939-9936-1_11.

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.

Keywords: Formalin-fixed, paraffin-embedded tissue; Immunohistochemistry; Western blot.

MeSH terms

  • Antigens, Differentiation, B-Lymphocyte / metabolism*
  • Biomarkers
  • Blotting, Western* / methods
  • Formaldehyde
  • Histocompatibility Antigens Class II / metabolism*
  • Immunohistochemistry* / methods
  • Paraffin Embedding
  • Protein Binding
  • Protein Transport
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism*
  • Tissue Fixation

Substances

  • Antigens, Differentiation, B-Lymphocyte
  • Biomarkers
  • Histocompatibility Antigens Class II
  • Receptors, Immunologic
  • invariant chain
  • macrophage migration inhibitory factor receptor
  • Formaldehyde