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liyan20220木虫 (著名写手)
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[求助]
求翻译一段中文为英文
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| 组装Nanodiscs前，首先将POPE和POPG胆酸钠配成80mM磷脂，160mM胆酸钠的母液，取476ul 0.2mM MSP蛋白母液，2.4ul POPG和胆酸钠母液，21.6ul POPE和胆酸钠母液，混匀后，补17.5ul 200mM 胆酸钠，因为POPE的相变温度是25℃，POPG的相变温度为-2℃，我们选用14℃和25℃对磷脂分子进行过夜孵育，孵育后加入疏水吸附剂Bio-Beads SM-2 (Bio-Rad， Hercules， CA)，室温孵育2h，自组装形成Nanodiscs。用注射器吸取组装后的Nanodiscs的上清，13000rpm,离心10min后，通过FPLC Superdex 200 16/60色谱层析柱(GE Healthcare)进一步纯化收集，流动相流速为1 ml/min。纯化的Nanodiscs样品的形貌和尺寸可以采用透射电镜进行检测。 |
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liyan20220: 金币+55, 翻译EPI+1, 谢谢你 2014-02-21 18:17:14
liyan20220: 金币+55, 翻译EPI+1, 谢谢你 2014-02-21 18:17:14
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Prior to Nanodiscs assembly , POPE and POPG were prepared for phospholipids 80mM and 160mM sodium cholate mother liquor, then 476ul 0.2mM MSP protein liquor, 2.4ul POPG and cholate liquor, 21.6ul POPE and sodium cholate liquor were added into .Blend the liquor and fill in 17.5ul 200mM sodium cholate.14 ℃ and 25 ℃ of phospholipid molecules overnight incubation were selected considering that POPE phase transition temperature is 25 ℃while POPG is -2 ℃ .Now put in hydrophobic adsorbent Bio-Beads SM-2 (Bio-Rad, Hercules, CA) and incubate at room temperature for 2h for the self-assemble of Nanodiscs. The supernatant were draw with a syringe for 13000rpm, 10min centrifugation and further purified by FPLC Superdex 200 16/60 chromatography column (GE Healthcare) with mobile phase flow rate of 1 ml / min. Transmission electron microscopy were used to detect the sample size and morphology of the Purified Nanodiscs., 自己再核对一下。 |
2楼2014-02-21 12:32:57
