An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis.

  title={An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis.},
  author={A. Mansour and P. A. Khalil and Noha A. Bejjani and R. Chatila and Carole Dagher-Hamalian and Wissam H. Faour},
  journal={Histology and histopathology},
  volume={32 3},
Deparaffinization of formalin-fixed paraffin embedded (FFPE) tissues with xylene currently remains a major challenge to the biomedical community. [...] Key Method A total of 79 different types of FFPE tissue sections of 8 µm thickness were obtained from various archived FFPE specimens. Deparaffinization was conducted by gently washing each section with around 1 ml of hot distilled water (≈80°C).Expand
3 Citations
Update on proteomic studies of formalin-fixed paraffin-embedded tissues
ABSTRACT Introduction: This review is an update on recent progress in proteomic studies of formalin-fixed paraffin-embedded (FFPE) tissues, which open the opportunity to investigate diseases andExpand
Molecular glycopathology by capillary electrophoresis: Analysis of the N‐glycome of formalin‐fixed paraffin‐embedded mouse tissue samples
Capillary electrophoresis with laser‐induced fluorescence (CE‐LIF) detection was used to analyze endoglycosidase released and fluorophore‐labeled N‐glycans from formalin‐fixed paraffin‐embeddedExpand
Developing a growing cancer profile database based on quantitative analysis of protein biomarkers in formalin-fixed paraffin-embedded specimens.
A continuously growing cancer patient profile database is proposed here to support "big data" profiling of these protein biomarkers alone or in combination, enabling next-generation retrospective-prospective analytics into the field of clinical diagnostics. Expand


Quantitative protein analysis from formalin‐fixed tissues: implications for translational clinical research and nanoscale molecular diagnosis
The receptor tyrosine kinase HER2, an important protein target for antibody based cancer treatment, was reliably measured in formalin‐fixed breast cancer biopsy samples when compared with measurement by immunohistochemistry and fluorescence in situ hybridization. Expand
A novel procedure for protein extraction from formalin‐fixed paraffin‐embedded tissues
A novel, short time‐consuming and cost‐effective method to extract full‐length, non‐degraded proteins from FFPE tissues and enough intact protein is obtained to be detected by Western blotting analysis. Expand
Successful Protein Extraction from Over-Fixed and Long-Term Stored Formalin-Fixed Tissues
Evaluating the protein extraction efficiency of FFPE tissues which had been formalin-fixed for up to 144 hours and tissue blocks that were stored for 20 years, comparing an established and a new commercial buffer system shows that the new buffer system is superior compared to the established one. Expand
Factors in Tissue Handling and Processing That Impact RNA Obtained From Formalin-fixed, Paraffin-embedded Tissue
It is determined that the middle region of gene suffers less damage by these processes as shown by real-time quantitative RT-PCR, which contributes to the establishment of objective standards for the processing and handling of tissue in surgical pathology. Expand
‘Tissue surrogates’ as a model for archival formalin-fixed paraffin-embedded tissues
Tissue surrogates formed from one or two proteins were used to evaluate extraction protocols for their ability to quantitatively extract proteins from the surrogates and revealed that the surrogate extracts contained a mixture of monomeric and multimeric proteins, regardless of the extraction protocol employed. Expand
Proteomic analysis of formalin-fixed paraffin-embedded tissue by MALDI imaging mass spectrometry
A protocol for analyzing protein in FFPE-TMAs using matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) for direct analysis of multiple tissue samples in a single experiment without extraction and purification of proteins is described. Expand
Development and validation of a novel protein extraction methodology for quantitation of protein expression in formalin‐fixed paraffin‐embedded tissues using western blotting
A novel heat‐induced antigen retrieval strategy using SDS‐containing Laemmli buffer for efficient intact protein recovery from formalin‐fixed tissues for subsequent analysis by western blotting is developed, and the protocol enables efficient extraction of non‐degraded, full‐length, immunoreactive protein. Expand
Enrichment of PrPSc in formalin-fixed, paraffin-embedded tissues prior to analysis by Western blot
  • E. Nicholson
  • Biology, Medicine
  • Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
  • 2011
A method of enriching PrPSc in formalin-fixed, paraffin-embedded tissues prior to Western blot analysis for the detection of PrP Sc is described, with 5 times the previously reported sample size used for analysis greatly enhancing the sensitivity of this procedure. Expand
Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections.
  • S. Shi, M. Key, K. Kalra
  • Chemistry, Medicine
  • The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
  • 1991
Good immunostaining results were obtained with a monoclonal antibody to vimentin as well as several different keratin antibodies on routine formalin-fixed tissue sections after pre-treatment of the slides with this method. Expand
A review of preanalytical factors affecting molecular, protein, and morphological analysis of formalin-fixed, paraffin-embedded (FFPE) tissue: how well do you know your FFPE specimen?
Based on the literature evidence, the molecular, proteomic, and morphological endpoints can be altered in formalin-fixed, paraffin-embedded specimens by suboptimal processing conditions. Expand