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【答案】应助回帖
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liyan20220: 金币+55, 翻译EPI+1, 谢谢你 2014-02-21 18:17:14
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.,
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