Global Protein Profiling in Processed Immunohistochemistry Tissue Sections.
SDS-SP3 protocol
formalin fixed and embedded brain sample
mass spectrometry
protein preparation
Journal
International journal of molecular sciences
ISSN: 1422-0067
Titre abrégé: Int J Mol Sci
Pays: Switzerland
ID NLM: 101092791
Informations de publication
Date de publication:
11 Jul 2023
11 Jul 2023
Historique:
received:
16
06
2023
revised:
07
07
2023
accepted:
08
07
2023
medline:
31
7
2023
pubmed:
29
7
2023
entrez:
29
7
2023
Statut:
epublish
Résumé
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.
Identifiants
pubmed: 37511068
pii: ijms241411308
doi: 10.3390/ijms241411308
pmc: PMC10379013
pii:
doi:
Substances chimiques
Formaldehyde
1HG84L3525
Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
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