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An effective placental cotyledons proteins extraction method for 2D gel electrophoresis

Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this s...

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Main Authors: Tan, Niu Jin, Jeffery Daim, Leona Daniela, Mohd Jamil, Amilia Afzan, Mohtarrudin, Norhafizah, Karuppiah, Thilakavathy
Format: Article
Language:English
Published: Wiley-VCH Verlag GmbH & Co. KGaA 2017
Online Access:http://psasir.upm.edu.my/id/eprint/59625/1/An%20effective%20placental%20cotyledons%20proteins%20extraction%20method%20for%202D%20gel%20electrophoresis.pdf
http://psasir.upm.edu.my/id/eprint/59625/
http://onlinelibrary.wiley.com/wol1/doi/10.1002/elps.201600377/abstract
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spelling my.upm.eprints.596252018-03-14T02:04:24Z http://psasir.upm.edu.my/id/eprint/59625/ An effective placental cotyledons proteins extraction method for 2D gel electrophoresis Tan, Niu Jin Jeffery Daim, Leona Daniela Mohd Jamil, Amilia Afzan Mohtarrudin, Norhafizah Karuppiah, Thilakavathy Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen. Wiley-VCH Verlag GmbH & Co. KGaA 2017 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/59625/1/An%20effective%20placental%20cotyledons%20proteins%20extraction%20method%20for%202D%20gel%20electrophoresis.pdf Tan, Niu Jin and Jeffery Daim, Leona Daniela and Mohd Jamil, Amilia Afzan and Mohtarrudin, Norhafizah and Karuppiah, Thilakavathy (2017) An effective placental cotyledons proteins extraction method for 2D gel electrophoresis. Electrophoresis, 38 (5). pp. 633-644. ISSN 0173-0835; ESSN: 1522-2683 http://onlinelibrary.wiley.com/wol1/doi/10.1002/elps.201600377/abstract 10.1002/elps.201600377
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen.
format Article
author Tan, Niu Jin
Jeffery Daim, Leona Daniela
Mohd Jamil, Amilia Afzan
Mohtarrudin, Norhafizah
Karuppiah, Thilakavathy
spellingShingle Tan, Niu Jin
Jeffery Daim, Leona Daniela
Mohd Jamil, Amilia Afzan
Mohtarrudin, Norhafizah
Karuppiah, Thilakavathy
An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
author_facet Tan, Niu Jin
Jeffery Daim, Leona Daniela
Mohd Jamil, Amilia Afzan
Mohtarrudin, Norhafizah
Karuppiah, Thilakavathy
author_sort Tan, Niu Jin
title An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
title_short An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
title_full An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
title_fullStr An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
title_full_unstemmed An effective placental cotyledons proteins extraction method for 2D gel electrophoresis
title_sort effective placental cotyledons proteins extraction method for 2d gel electrophoresis
publisher Wiley-VCH Verlag GmbH & Co. KGaA
publishDate 2017
url http://psasir.upm.edu.my/id/eprint/59625/1/An%20effective%20placental%20cotyledons%20proteins%20extraction%20method%20for%202D%20gel%20electrophoresis.pdf
http://psasir.upm.edu.my/id/eprint/59625/
http://onlinelibrary.wiley.com/wol1/doi/10.1002/elps.201600377/abstract
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score 13.211869

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