An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees
Proteomic analysis of plants relies on high yields of pure protein. In plants, protein extraction and purification present a great challenge due to accumulation of a large amount of interfering substances, including polysaccharides, polyphenols, and secondary metabolites. Therefore, it is necessary...
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International Union of Biochemistry and Molecular Biology
2013
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my.upm.eprints.281422017-09-28T07:58:08Z http://psasir.upm.edu.my/id/eprint/28142/ An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees Talei, Daryush Valdiani, Alireza Abdullah, Mohd Puad Proteomic analysis of plants relies on high yields of pure protein. In plants, protein extraction and purification present a great challenge due to accumulation of a large amount of interfering substances, including polysaccharides, polyphenols, and secondary metabolites. Therefore, it is necessary to modify the extraction protocols. A study was conducted to compare four protein extraction and precipitation methods for proteomic analysis. The results showed significant differences in protein content among the four methods. The chloroform-trichloroacetic acid-acetone method using 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer provided the best results in terms of protein content, pellets, spot resolution, and intensity of unique spots detected. An overall of 83 qualitative or quantitative significant differential spots were found among the four methods. Based on the 2-DE gel map, the method is expected to benefit the development of high-level proteomic and biochemical studies of Andrographis paniculata, which may also be applied to other recalcitrant medicinal plant tissues. International Union of Biochemistry and Molecular Biology 2013 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/28142/1/An%20effective%20protein%20extraction%20method%20for%20two.pdf Talei, Daryush and Valdiani, Alireza and Abdullah, Mohd Puad (2013) An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees. Biotechnology and Applied Biochemistry, 60 (5). pp. 521-526. ISSN 0885-4513; ESSN: 1470-8744 10.1002/bab.1126 |
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Proteomic analysis of plants relies on high yields of pure protein. In plants, protein extraction and purification present a great challenge due to accumulation of a large amount of interfering substances, including polysaccharides, polyphenols, and secondary metabolites. Therefore, it is necessary to modify the extraction protocols. A study was conducted to compare four protein extraction and precipitation methods for proteomic analysis. The results showed significant differences in protein content among the four methods. The chloroform-trichloroacetic acid-acetone method using 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer provided the best results in terms of protein content, pellets, spot resolution, and intensity of unique spots detected. An overall of 83 qualitative or quantitative significant differential spots were found among the four methods. Based on the 2-DE gel map, the method is expected to benefit the development of high-level proteomic and biochemical studies of Andrographis paniculata, which may also be applied to other recalcitrant medicinal plant tissues. |
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Talei, Daryush Valdiani, Alireza Abdullah, Mohd Puad |
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Talei, Daryush Valdiani, Alireza Abdullah, Mohd Puad An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
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Talei, Daryush Valdiani, Alireza Abdullah, Mohd Puad |
author_sort |
Talei, Daryush |
title |
An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
title_short |
An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
title_full |
An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
title_fullStr |
An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
title_full_unstemmed |
An effective protein extraction method for two-dimensional electrophoresis in the anticancer herb Andrographis paniculata Nees |
title_sort |
effective protein extraction method for two-dimensional electrophoresis in the anticancer herb andrographis paniculata nees |
publisher |
International Union of Biochemistry and Molecular Biology |
publishDate |
2013 |
url |
http://psasir.upm.edu.my/id/eprint/28142/1/An%20effective%20protein%20extraction%20method%20for%20two.pdf http://psasir.upm.edu.my/id/eprint/28142/ |
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