Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates
Abstract An efficient protein extraction methodology is quite important for sample preparation and subsequent 2‐D PAGE and MS analysis. Cell lysis is the first step in protein extraction and purification. Many techniques are available for cell disruption, including physical and detergent‐based metho...
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crwiley:10.1002/elps.200700838 2024-06-02T07:58:29+00:00 Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates Fulgentini, Lorenzo Marangoni, Roberto Colombetti, Giuliano 2008 http://dx.doi.org/10.1002/elps.200700838 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felps.200700838 https://onlinelibrary.wiley.com/doi/full/10.1002/elps.200700838 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor ELECTROPHORESIS volume 29, issue 11, page 2411-2412 ISSN 0173-0835 1522-2683 journal-article 2008 crwiley https://doi.org/10.1002/elps.200700838 2024-05-03T11:05:05Z Abstract An efficient protein extraction methodology is quite important for sample preparation and subsequent 2‐D PAGE and MS analysis. Cell lysis is the first step in protein extraction and purification. Many techniques are available for cell disruption, including physical and detergent‐based methods. Here, we report on a very fast and efficient detergent‐free Tris‐based method to extract the soluble fraction proteins of extremophile ciliates, comparing it with a detergent‐based protocol. This comparison has been carried out by means of 2‐D PAGE and subsequent MALDI‐compatible silver staining of protein samples obtained from the intensely pigmented hypersaline ciliate Fabrea salina and the Antarctic hypotrich ciliate Euplotes focardii . Our results indicate that this fast and easy extraction method allows to obtain more clear crude extracts and more spot‐abundant polyacrylamide gels. Article in Journal/Newspaper Antarc* Antarctic Wiley Online Library Antarctic The Antarctic ELECTROPHORESIS 29 11 2411 2412 |
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English |
description |
Abstract An efficient protein extraction methodology is quite important for sample preparation and subsequent 2‐D PAGE and MS analysis. Cell lysis is the first step in protein extraction and purification. Many techniques are available for cell disruption, including physical and detergent‐based methods. Here, we report on a very fast and efficient detergent‐free Tris‐based method to extract the soluble fraction proteins of extremophile ciliates, comparing it with a detergent‐based protocol. This comparison has been carried out by means of 2‐D PAGE and subsequent MALDI‐compatible silver staining of protein samples obtained from the intensely pigmented hypersaline ciliate Fabrea salina and the Antarctic hypotrich ciliate Euplotes focardii . Our results indicate that this fast and easy extraction method allows to obtain more clear crude extracts and more spot‐abundant polyacrylamide gels. |
format |
Article in Journal/Newspaper |
author |
Fulgentini, Lorenzo Marangoni, Roberto Colombetti, Giuliano |
spellingShingle |
Fulgentini, Lorenzo Marangoni, Roberto Colombetti, Giuliano Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
author_facet |
Fulgentini, Lorenzo Marangoni, Roberto Colombetti, Giuliano |
author_sort |
Fulgentini, Lorenzo |
title |
Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
title_short |
Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
title_full |
Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
title_fullStr |
Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
title_full_unstemmed |
Optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
title_sort |
optimizing soluble protein extraction and two‐dimensional polyacrylamide gel electrophoresis quality for extremophile ciliates |
publisher |
Wiley |
publishDate |
2008 |
url |
http://dx.doi.org/10.1002/elps.200700838 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Felps.200700838 https://onlinelibrary.wiley.com/doi/full/10.1002/elps.200700838 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
ELECTROPHORESIS volume 29, issue 11, page 2411-2412 ISSN 0173-0835 1522-2683 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/elps.200700838 |
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ELECTROPHORESIS |
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29 |
container_issue |
11 |
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2411 |
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2412 |
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1800741854070702080 |