An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis
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...
محفوظ في:
| المؤلف الرئيسي: | |
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| مؤلفون آخرون: | , , , , |
| التنسيق: | article |
| منشور في: |
2017
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| الوصول للمادة أونلاين: | http://hdl.handle.net/10725/10662 http://dx.doi.org/10.14670/HH-11-789 http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://pubmed.ncbi.nlm.nih.gov/27264533/ |
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| _version_ | 1864513487056666624 |
|---|---|
| author | Mansour, Anthony G. |
| author2 | Abou-Khalil, Pamela Bejjani, Noha Chatila, Rajaa Dagher-Hamalian, Carol Faour, Wissam H. |
| author2_role | author author author author author |
| author_facet | Mansour, Anthony G. Abou-Khalil, Pamela Bejjani, Noha Chatila, Rajaa Dagher-Hamalian, Carol Faour, Wissam H. |
| author_role | author |
| dc.creator.none.fl_str_mv | Mansour, Anthony G. Abou-Khalil, Pamela Bejjani, Noha Chatila, Rajaa Dagher-Hamalian, Carol Faour, Wissam H. |
| dc.date.none.fl_str_mv | 2017 2019-05-23T12:08:47Z 2019-05-23T12:08:47Z 2019-05-23 |
| dc.identifier.none.fl_str_mv | 1699-5848 http://hdl.handle.net/10725/10662 http://dx.doi.org/10.14670/HH-11-789 Mansour, A. G., Khalil, P. A., Bejjani, N., Chatila, R., Dagher-Hamalian, C., & Faour, W. H. (2016). 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. http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://pubmed.ncbi.nlm.nih.gov/27264533/ |
| dc.language.none.fl_str_mv | en |
| dc.relation.none.fl_str_mv | Histology and Histopathology |
| dc.rights.*.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.title.none.fl_str_mv | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| dc.type.none.fl_str_mv | Article info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
| description | 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. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | LAURepo_b7f6170c013cc5cf93a6edf9e4a2b4c1 |
| identifier_str_mv | 1699-5848 Mansour, A. G., Khalil, P. A., Bejjani, N., Chatila, R., Dagher-Hamalian, C., & Faour, W. H. (2016). 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. |
| language_invalid_str_mv | en |
| network_acronym_str | LAURepo |
| network_name_str | Lebanese American University repository |
| oai_identifier_str | oai:laur.lau.edu.lb:10725/10662 |
| publishDate | 2017 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| spelling | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysisMansour, Anthony G.Abou-Khalil, PamelaBejjani, NohaChatila, RajaaDagher-Hamalian, CarolFaour, Wissam H.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.PublishedN/A2019-05-23T12:08:47Z2019-05-23T12:08:47Z20172019-05-23Articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1699-5848http://hdl.handle.net/10725/10662http://dx.doi.org/10.14670/HH-11-789Mansour, A. G., Khalil, P. A., Bejjani, N., Chatila, R., Dagher-Hamalian, C., & Faour, W. H. (2016). 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.http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.phphttps://pubmed.ncbi.nlm.nih.gov/27264533/enHistology and Histopathologyinfo:eu-repo/semantics/openAccessoai:laur.lau.edu.lb:10725/106622024-05-20T06:46:30Z |
| spellingShingle | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis Mansour, Anthony G. |
| status_str | publishedVersion |
| title | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| title_full | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| title_fullStr | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| title_full_unstemmed | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| title_short | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| title_sort | An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis |
| url | http://hdl.handle.net/10725/10662 http://dx.doi.org/10.14670/HH-11-789 http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://pubmed.ncbi.nlm.nih.gov/27264533/ |