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chensang

新虫 (初入文坛)

[求助] 蛋白沉淀 已有1人参与

求助,TCA丙酮沉淀蛋白的具体步骤

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zhouxfd

木虫 (正式写手)

【答案】应助回帖

感谢参与,应助指数 +1
Acetone precipitation
1.        Add same volume of 20% TCA (in water).
2.        Place on ice for at least 1 hour.
3.        Add 4 volume of 100% cold acetone (-20℃) into your protein solution.
4.        Mix well and put it in -20℃ freezer overnight (or -80℃ freezer for at least 1 hour)
5.        Collect precipitate by centrifugation (20min, max speed-at least 20,000 × g, 4C).
6.        Wash the pellet with 1 ml of ice-cold 80% acetone (-20℃ freezer, overnight).
7.        Repeat step 4 twice [total thrice wash].
        Completely resuspend the pellet each time with vigorous vortex.
        Do not overdry it out. I usually place proteins for 7 min 30 sec. with the -20℃ block.
2楼2017-07-19 00:44:45
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chensang

新虫 (初入文坛)

引用回帖:
2楼: Originally posted by zhouxfd at 2017-07-19 00:44:45
Acetone precipitation
1.        Add same volume of 20% TCA (in water).
2.        Place on ice for at least 1 hour.
3.        Add 4 volume of 100% cold acetone (-20℃) into your protein solution.
4.        Mix well and put i ...

谢谢

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3楼2017-07-19 23:56:42
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iophmx

新虫 (初入文坛)

请问楼主这个实验你做了吗?负20度冷丙酮你用什么容器放冰箱的啊

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4楼2017-12-01 11:06:18
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