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349592850

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[求助] 怎样避免蛋白交联呢?

请问一下,我用巯基乙胺去修饰蛋白,蛋白的巯基发生了交联,除了通过在无氧的条件下进行反应可以避免蛋白交联,还有什么别的方法吗?另:修饰后的蛋白上的巯基后期还会用到!谢谢大家!
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aircraftvip

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349592850: 金币+5, ★★★很有帮助, 十分感谢!十分感谢! 2013-06-04 08:33:04
红舟流星: 金币+1, 专家考核, 欢迎来生物材料板块应助交流 2013-06-04 23:36:25
you could also consider to add reducing agent into your working solution, such as TCEP.
besides, lower pH value(3-4) would lower the possibility of disulfide recombination due to protonation of sulfhydryl groups.
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2楼2013-06-04 01:09:29
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349592850

铁杆木虫 (正式写手)

引用回帖:
2楼: Originally posted by aircraftvip at 2013-06-04 01:09:29
you could also consider to add reducing agent into your working solution, such as TCEP.
besides, lower pH value(3-4) would lower the possibility of disulfide recombination due to protonation of s ...

十分感谢!我想再请教一下,利用TCEP阻止蛋白的巯基交联,那修饰后的蛋白上的巯基还能否再发生交联呢?比如利用氧化剂如双氧水之类的还可以使其交联吗?
3楼2013-06-04 08:27:13
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aircraftvip

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349592850: 金币+5, ★★★很有帮助, Thank you very much! I may have a try! Thank you! 2013-06-04 20:50:02
引用回帖:
3楼: Originally posted by 349592850 at 2013-06-04 08:27:13
十分感谢!我想再请教一下,利用TCEP阻止蛋白的巯基交联,那修饰后的蛋白上的巯基还能否再发生交联呢?比如利用氧化剂如双氧水之类的还可以使其交联吗?...

TCEP can even catalyze certain reactions. If you rely on oxidation, -SH groups can be reused crosslinking when H2O2 is more than TCEP.
But be careful with the possible toxicity of TCEP.
格物致知,知行合一
4楼2013-06-04 17:49:00
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