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海洋之滨

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[求助] DTT会打开金硫键吗?

实验中用到含DTT的一个蛋白,蛋白溶液在金电极上反应一段时间后,阻抗明显减小,电极上只有巯基DNA和巯基己烷,确定是因为DTT的存在。不知道是因为DTT打开了金硫键还是因为二硫键的存在,怎么去除DTT或者怎么消除影响?

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引用回帖:
3楼: Originally posted by aircraftvip at 2012-12-11 23:17:17
Or consider desalting, but it is much more laborious.
``Dimerization greatly lowers the efficiency of subsequent coupling reactions such as DNA immobilization on gold in biosensors. Typically DTT i ...

怎么不去http://muchong.com/bbs/viewthread.php?tid=4945856&page=8&fpage=领取十一月季军奖励啊?
红楼一梦终难寻扁舟载酒且慢行落花流水依惜别月伴明星照前路
4楼2012-12-12 00:15:47
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aircraftvip

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【答案】应助回帖

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感谢参与,应助指数 +1
红舟流星: 金币+1, 专家考核, 欢迎来到生物功能材料板块应助交流 2012-12-12 00:15:13
海洋之滨: 金币+10, ★有帮助 2012-12-12 10:17:52
DTT自身带的两个巯基也可以跟金电极络合。
建议楼主看看这篇文献,
The Structure of Dithiothreitol Monolayers on Au(111)
http://bolt.lakeheadu.ca/~physww ... surface_grp/DTT.pdf

排除DTT的干扰,建议你用不含巯基的还原剂,比如硼氢化钠,TCEP
格物致知,知行合一
2楼2012-12-11 23:13:27
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aircraftvip

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海洋之滨: 金币+20, ★有帮助 2012-12-12 10:18:01
Or consider desalting, but it is much more laborious.
``Dimerization greatly lowers the efficiency of subsequent coupling reactions such as DNA immobilization on gold in biosensors. Typically DTT is mixed with a DNA solution and allowed to react, and then is removed by filtration (for the solid catalyst) or by chromatography (for the liquid form). The DTT removal procedure is often called "desalting."``
---Wikipedia
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3楼2012-12-11 23:17:17
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海洋之滨

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引用回帖:
2楼: Originally posted by aircraftvip at 2012-12-11 23:13:27
DTT自身带的两个巯基也可以跟金电极络合。
建议楼主看看这篇文献,
The Structure of Dithiothreitol Monolayers on Au(111)
http://bolt.lakeheadu.ca/~physwww/facNstaff/gallagher/surface_grp/DTT.pdf

排除 ...

有可能DTT上的巯基取代了原来的DNA和封闭剂或者原来的-SH因为形成二硫键而二硫键也能与金作用,DTT又恰好能打断二硫键。但实验过程中因为DTT的存在几乎能把修饰电极变成裸电极,所以还是很疑惑。 另外,DTT本身是还原剂,是不是需要氧化剂来消除???
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5楼2012-12-12 10:24:35
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