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HERO ID
1104477
Reference Type
Journal Article
Title
Evaluation of ethanol-fixed, paraffin-embedded tissues for proteomic applications
Author(s)
Ahram, M; Flaig, MJ; Gillespie, JW; Duray, PH; Linehan, WM; Ornstein, DK; Niu, S; Zhao, Y; Petricoin, EF; Emmert-Buck, MR
Year
2003
Is Peer Reviewed?
1
Journal
Proteomics
ISSN:
1615-9853
EISSN:
1615-9861
Volume
3
Issue
4
Page Numbers
413-421
Language
English
PMID
12687609
DOI
10.1002/pmic.200390056
Abstract
We previously reported that ethanol fixation and paraffin embedding of tissues produce excellent histomorphology and good preservation of macromolecules. Here, we present a detailed evaluation of ethanol-fixed tissues for proteomic initiatives. When proteins were extracted from ethanol-fixed, paraffin-embedded prostate tissue, resolved by two-dimensional gel electrophoresis (2-DE), and stained by standard methods, several hundred protein molecules could be detected and successfully analyzed by mass spectrometry. Protein profiles obtained from ethanol-fixed tissues were highly similar to those observed from frozen tissues, in contrast to the poor protein recovery from formalin-fixed material. The protein content of specific cells that were microdissected from ethanol-fixed tissue sections using laser capture microdissection could also be successfully analyzed by 2-DE. We observed that eosin staining of tissue sections had a detrimental effect on protein separation, whereas hematoxylin staining had minimal consequence. In order to illustrate the applicability of ethanol-fixed tissues for proteomic discovery studies, we compared the protein profiles of patient-matched, normal prostatic epithelial cells and invasive adenocarcinoma cells obtained from ethanol-fixed, paraffin-embedded tissues. A number of differentially expressed proteins was discovered and identified by mass spectrometry. Immunohistochemical analyses performed on ethanol-fixed tissue sections were in agreement with the proteomic discovery findings. In light of these results, we conclude that ethanol-fixed tissues can be successfully utilized for proteomic analyses.
Tags
IRIS
•
Methanol (Non-Cancer)
Ramazzini
Other
•
Ramazzini Institute
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