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Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks

BACKGROUND: Methanol fixed and paraffin embedded (MFPE) cellblocks are an essential cytology preparation. However, MFPE cellblocks often contain limited material and their relatively small size has caused them to be overlooked in biomarker discovery. Advances in the field of molecular biotechnology...

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Autores principales: Kokkat, Theresa J., McGarvey, Diane, Patel, Miral S., Tieniber, Andrew D., LiVolsi, Virginia A., Baloch, Zubair W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications & Media Pvt Ltd 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3869970/
https://www.ncbi.nlm.nih.gov/pubmed/24403950
http://dx.doi.org/10.4103/1742-6413.122299
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author Kokkat, Theresa J.
McGarvey, Diane
Patel, Miral S.
Tieniber, Andrew D.
LiVolsi, Virginia A.
Baloch, Zubair W.
author_facet Kokkat, Theresa J.
McGarvey, Diane
Patel, Miral S.
Tieniber, Andrew D.
LiVolsi, Virginia A.
Baloch, Zubair W.
author_sort Kokkat, Theresa J.
collection PubMed
description BACKGROUND: Methanol fixed and paraffin embedded (MFPE) cellblocks are an essential cytology preparation. However, MFPE cellblocks often contain limited material and their relatively small size has caused them to be overlooked in biomarker discovery. Advances in the field of molecular biotechnology have made it possible to extract proteins from formalin fixed and paraffin embedded (FFPE) tissue blocks. In contrast, there are no established methods for extracting proteins from MFPE cellblocks. We investigated commonly available CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) buffer, as well as two commercially available Qiagen(®) kits and compared their effectiveness on MFPE tissue for protein yields. MATERIALS AND METHODS: MFPE blocks were made by Cellient™ automated system using human tissue specimens from normal and malignant specimens collected in ThinPrep™ Vials. Protein was extracted from Cellient-methanol fixed and paraffin embedded blocks with CHAPS buffer method as well as FFPE and Mammalian Qiagen(®) kits. RESULTS: Comparison of protein yields demonstrated the effectiveness of various protein extraction methods on MFPE cellblocks. CONCLUSION: In the current era of minimally invasive techniques to obtain minimal amount of tissue for diagnostic and prognostic purposes, the use of commercial and lab made buffer on low weight MFPE scrapings obtained by Cellient(®) processor opens new possibilities for protein biomarker research.
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spelling pubmed-38699702014-01-08 Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks Kokkat, Theresa J. McGarvey, Diane Patel, Miral S. Tieniber, Andrew D. LiVolsi, Virginia A. Baloch, Zubair W. Cytojournal Methodology Articles BACKGROUND: Methanol fixed and paraffin embedded (MFPE) cellblocks are an essential cytology preparation. However, MFPE cellblocks often contain limited material and their relatively small size has caused them to be overlooked in biomarker discovery. Advances in the field of molecular biotechnology have made it possible to extract proteins from formalin fixed and paraffin embedded (FFPE) tissue blocks. In contrast, there are no established methods for extracting proteins from MFPE cellblocks. We investigated commonly available CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) buffer, as well as two commercially available Qiagen(®) kits and compared their effectiveness on MFPE tissue for protein yields. MATERIALS AND METHODS: MFPE blocks were made by Cellient™ automated system using human tissue specimens from normal and malignant specimens collected in ThinPrep™ Vials. Protein was extracted from Cellient-methanol fixed and paraffin embedded blocks with CHAPS buffer method as well as FFPE and Mammalian Qiagen(®) kits. RESULTS: Comparison of protein yields demonstrated the effectiveness of various protein extraction methods on MFPE cellblocks. CONCLUSION: In the current era of minimally invasive techniques to obtain minimal amount of tissue for diagnostic and prognostic purposes, the use of commercial and lab made buffer on low weight MFPE scrapings obtained by Cellient(®) processor opens new possibilities for protein biomarker research. Medknow Publications & Media Pvt Ltd 2013-11-28 /pmc/articles/PMC3869970/ /pubmed/24403950 http://dx.doi.org/10.4103/1742-6413.122299 Text en Copyright: © 2013 Kokkat, et al.; licensee Cytopathology Foundation Inc. http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Articles
Kokkat, Theresa J.
McGarvey, Diane
Patel, Miral S.
Tieniber, Andrew D.
LiVolsi, Virginia A.
Baloch, Zubair W.
Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title_full Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title_fullStr Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title_full_unstemmed Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title_short Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
title_sort protein extraction from methanol fixed paraffin embedded tissue blocks: a new possibility using cell blocks
topic Methodology Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3869970/
https://www.ncbi.nlm.nih.gov/pubmed/24403950
http://dx.doi.org/10.4103/1742-6413.122299
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