Global Protein Profiling in Processed Immunohistochemistry Tissue Sections

Int J Mol Sci. 2023 Jul 11;24(14):11308. doi: 10.3390/ijms241411308.


Tissue sections, which are widely used in research and diagnostic laboratories and have already been examined by immunohistochemistry (IHC), may subsequently provide a resource for proteomic studies, even though only small amount of protein is available. Therefore, we established a workflow for tandem mass spectrometry-based protein profiling of IHC specimens and characterized defined brain area sections. We investigated the CA1 region of the hippocampus dissected from brain slices of adult C57BL/6J mice. The workflow contains detailed information on sample preparation from brain slices, including removal of antibodies and cover matrices, dissection of region(s) of interest, protein extraction and digestion, mass spectrometry measurement, and data analysis. The Gene Ontology (GO) knowledge base was used for further annotation. Literature searches and Gene Ontology annotation of the detected proteins verify the applicability of this method for global protein profiling using formalin-fixed and embedded material and previously used IHC slides.

Keywords: SDS-SP3 protocol; formalin fixed and embedded brain sample; mass spectrometry; protein preparation.

MeSH terms

  • Animals
  • Formaldehyde* / chemistry
  • Immunohistochemistry
  • Mice
  • Mice, Inbred C57BL
  • Paraffin Embedding
  • Proteins / analysis
  • Proteomics* / methods
  • Tandem Mass Spectrometry
  • Tissue Fixation / methods


  • Formaldehyde
  • Proteins

Grants and funding

This research received no external funding.