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2-D PAGE and MS analysis of proteins from formalin-fixed, paraffin-embedded tissues
- Source :
- PROTEOMICS. 9:4329-4339
- Publication Year :
- 2009
- Publisher :
- Wiley, 2009.
-
Abstract
- In the past decade, encouraging results have been obtained in extraction and analysis of proteins from formalin-fixed, paraffin-embedded (FFPE) tissues. However, 2-D PAGE protein maps with satisfactory proteomic information and comparability to fresh tissues have never been described to date. In the present study, we report 2-D PAGE separation and MS identification of full-length proteins extracted from FFPE skeletal muscle tissue. The 2-D protein profiles obtained from FFPE tissues could be matched to those achieved from frozen tissues replicates. Up to 250 spots were clearly detected in 2-D maps of proteins from FFPE tissue following standard mass-compatible silver staining. Protein spots from both FFPE and frozen tissue 2-D gels were excised, subjected to in situ hydrolysis, and identified by MS analysis. Matched spots produced matched protein identifications. Moreover, 2-D protein maps from FFPE tissues were successfully subjected to Western immunoblotting, producing comparable results to fresh-frozen tissues. In conclusion, this study provides evidence that, when adequately extracted, full-length proteins from FFPE tissues might be suitable to 2-D PAGE-MS analysis, allowing differential proteomic studies on the vast existing archives of healthy and pathological-fixed tissues.
- Subjects :
- Silver Staining
Formalin fixed paraffin embedded
Immunoblotting
Muscle Proteins
Biology
Proteomics
Biochemistry
Mass Spectrometry
Specimen Handling
Silver stain
Formaldehyde
Skeletal Muscle Tissue
Animals
Electrophoresis, Gel, Two-Dimensional
Isoelectric Point
Frozen tissue
Muscle, Skeletal
Molecular Biology
Western immunoblotting
Paraffin Embedding
Sheep
2 d page
Hydrolysis
Ms analysis
Proteins
Molecular biology
Subjects
Details
- ISSN :
- 16159853
- Volume :
- 9
- Database :
- OpenAIRE
- Journal :
- PROTEOMICS
- Accession number :
- edsair.doi.dedup.....213c5304b69afaef105ea0604d9efb7e