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liyan20220木蟲 (著名寫手)
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[求助]
求翻譯一段中文為英文
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| 組裝Nanodiscs前，首先將POPE和POPG膽酸鈉配成80mM磷脂，160mM膽酸鈉的母液，取476ul 0.2mM MSP蛋白母液，2.4ul POPG和膽酸鈉母液，21.6ul POPE和膽酸鈉母液，混勻后，補(bǔ)17.5ul 200mM 膽酸鈉，因?yàn)镻OPE的相變溫度是25℃，POPG的相變溫度為-2℃，我們選用14℃和25℃對(duì)磷脂分子進(jìn)行過夜孵育，孵育后加入疏水吸附劑Bio-Beads SM-2 (Bio-Rad， Hercules， CA)，室溫孵育2h，自組裝形成Nanodiscs。用注射器吸取組裝后的Nanodiscs的上清，13000rpm,離心10min后，通過FPLC Superdex 200 16/60色譜層析柱(GE Healthcare)進(jìn)一步純化收集，流動(dòng)相流速為1 ml/min。純化的Nanodiscs樣品的形貌和尺寸可以采用透射電鏡進(jìn)行檢測(cè)。 |
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【答案】應(yīng)助回帖
★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ...
liyan20220: 金幣+55, 翻譯EPI+1, 謝謝你 2014-02-21 18:17:14
liyan20220: 金幣+55, 翻譯EPI+1, 謝謝你 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., 自己再核對(duì)一下。 |
2樓2014-02-21 12:32:57
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