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Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue
Published on: September 2, 2013
Proteomic analysis of PAXgene-fixed tissues
Bilge Ergin1, Stephan Meding, Rupert Langer
1Institute of Pathology, Technische Universität München, Munich, Germany.
Journal of Proteome Research
|September 4, 2010
Summary
A new formalin-free fixative, PAXgene, effectively preserves proteins in paraffin-embedded tissues. This method enables high-quality protein extraction for proteomic analysis, comparable to frozen samples.
Area of Science:
- Biochemistry
- Molecular Biology
- Pathology
Background:
- Formalin fixation and paraffin embedding (FFPE) is standard for tissue preservation but challenges proteomic analysis due to protein cross-linking.
- Existing methods limit the utility of FFPE tissues for advanced molecular and proteomic studies.
Purpose of the Study:
- To evaluate PAXgene, a novel formalin-free fixative, as an alternative for tissue preservation.
- To demonstrate the suitability of PAXgene-fixed, paraffin-embedded (PFPE) tissues for proteomic applications.
Main Methods:
- PAXgene fixation of various human and mouse tissues.
- Protein extraction and quantification.
- Western blot analysis for protein detection (membrane, cytoplasmic, nuclear, phosphorylated proteins).
- Matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry.
Main Results:
- High protein yields from PFPE tissues comparable to frozen samples.
- Western blot analysis showed comparable protein detection in PFPE tissues to cryopreserved samples.
- PFPE tissue protein distribution is suitable for MALDI imaging mass spectrometry.
Conclusions:
- PAXgene is a viable formalin-free alternative for tissue fixation.
- PFPE tissues support extensive protein biomarker studies and multimodal proteomic analysis.
- PAXgene offers potential as a novel fixative for modern pathology.
