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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\u2013trichloroacetic acid\u2013acetone method using 4\u2010(2\u2010hydroxyethyl)\u20101\u2010piperazineethanesulfonic acid (<jats:styled-content style=\"fixed-case\">HEPES<\/jats:styled-content>) 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\u2010<jats:styled-content style=\"fixed-case\">DE<\/jats:styled-content> gel map, the method is expected to benefit the development of high\u2010level proteomic and biochemical studies of <jats:styled-content style=\"fixed-case\"><jats:italic>A<\/jats:italic><\/jats:styled-content><jats:italic>ndrographis paniculata<\/jats:italic>, which may also be applied to other recalcitrant medicinal plant tissues.<\/jats:p>","DOI":"10.1002\/bab.1126","type":"journal-article","created":{"date-parts":[[2013,5,31]],"date-time":"2013-05-31T16:13:18Z","timestamp":1370016798000},"page":"521-526","source":"Crossref","is-referenced-by-count":6,"title":["An effective protein extraction method for two\u2010dimensional electrophoresis in the anticancer herb <scp><i>A<\/i><\/scp><i>ndrographis paniculata<\/i>\u00a0<scp>N<\/scp>ees."],"prefix":"10.1002","volume":"60","author":[{"given":"Daryush","family":"Talei","sequence":"first","affiliation":[{"name":"Medicinal Plant Research Center Shahed University  Tehran Iran"},{"name":"Department of Cell and Molecular Biology Faculty of Biotechnology and Biomolecular Sciences Universiti Putra Malaysia  Selangor DE Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alireza","family":"Valdiani","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Faculty of Biotechnology and Biomolecular Sciences Universiti Putra Malaysia  Selangor DE Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mohd Abdullah","family":"Puad","sequence":"additional","affiliation":[{"name":"Department of Cell and Molecular Biology Faculty of Biotechnology and Biomolecular Sciences Universiti Putra Malaysia  Selangor DE Malaysia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,9,19]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1079\/IVP2003520"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-011-1341-x"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1248\/cpb.42.1216"},{"key":"e_1_2_8_5_1","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1076\/1388-0209(200007)3831-SFT204","volume":"38","author":"Rajani M.","year":"2000","journal-title":"Pharm. 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