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

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dc.contributor.author Mansour, Anthony G.
dc.contributor.author Abou-Khalil, Pamela
dc.contributor.author Bejjani, Noha
dc.contributor.author Chatila, Rajaa
dc.contributor.author Dagher-Hamalian, Carol
dc.contributor.author Faour, Wissam H.
dc.date.accessioned 2019-05-23T12:08:47Z
dc.date.available 2019-05-23T12:08:47Z
dc.date.copyright 2017 en_US
dc.date.issued 2019-05-23
dc.identifier.issn 1699-5848 en_US
dc.identifier.uri http://hdl.handle.net/10725/10662
dc.description.abstract Deparaffinization of formalin-fixed paraffin embedded (FFPE) tissues with xylene currently remains a major challenge to the biomedical community. We developed an efficient xylene-free protocol to isolate proteins from archived FFPE human tissue sections. 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). The deparaffinized tissues were homogenized in lysis buffer, and the isolated proteins were quantified and efficiently resolved using western blot analysis for the presence of Protein kinase B (PKB/AKT) and β-actin. Moreover, a significant amount of proteins was successfully isolated with an average of 2.31 µg/µl. The migration pattern of AKT and β-actin obtained from the specimens was similar to the positive control obtained from protein lysates prepared from in vitro cultured MDA231 cancer cell lines. AKT was successfully identified in all specimens, and β-actin protein was resolved with an efficiency higher than 80%. The entire extraction procedure requires only 20 minutes. This newly developed technique is an efficient, safe, cost-effective, and rapid method to isolate proteins from FFPE tissue sections adequate for molecular analysis en_US
dc.language.iso en en_US
dc.title An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis en_US
dc.type Article en_US
dc.description.version Published en_US
dc.author.school SOM en_US
dc.author.idnumber 200902770 en_US
dc.author.idnumber 200904962 en_US
dc.author.idnumber 200902750 en_US
dc.author.department N/A en_US
dc.description.embargo N/A en_US
dc.relation.journal Histology and Histopathology en_US
dc.journal.volume 2017 en_US
dc.journal.issue 32 en_US
dc.article.pages 307-313 en_US
dc.keywords Pathology en_US
dc.keywords Cancer en_US
dc.keywords Paraffin en_US
dc.keywords Western blot en_US
dc.keywords Protein isolation, en_US
dc.keywords Xylene en_US
dc.identifier.doi http://dx.doi.org/10.14670/HH-11-789 en_US
dc.identifier.ctation Mansour, A. G., Khalil, P. A., Bejjani, N., Chatila, R., Dagher-Hamalian, C., & Faour, W. H. (2017). An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis. Histology and histopathology, 32(3), 307-313. en_US
dc.author.email carole.hamalian@lau.edu.lb en_US
dc.author.email wissam.faour@lau.edu.lb en_US
dc.author.email rajaa.chatila@lau.edu.lb en_US
dc.identifier.tou http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php en_US
dc.identifier.url https://pdfs.semanticscholar.org/37d2/e5e30540ffc7bd867bc5bedc2747fd1d5bbb.pdf en_US
dc.orcid.id https://orcid.org/0000-0002-0120-2275 en_US
dc.author.affiliation Lebanese American University en_US


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