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岁月微醺

金虫 (小有名气)

[交流] 请问有做细胞浆/核蛋白提取的朋友吗?求指导分析

最近在做这个,都是按照说明书做的,可是蛋白含量一直很低,而且曝光检测不到目的条带,请问是怎么回事啊?该如何改正?
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hc-material

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给的信息太少,不好分析。
2楼2015-08-25 17:15:23
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gJordys

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岁月微醺: 金币+5 2015-08-27 12:46:14
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3楼2015-08-25 19:42:29
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gJordys

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4楼2015-08-25 19:43:32
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gJordys

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5楼2015-08-25 19:45:32
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fuyuandj86

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是核蛋白浓度低吗?核蛋白浓度低很正常。一般分离以后细胞质蛋白是细胞核蛋白总量的5-10倍。
你用的核蛋白的内参是什么?核蛋白不能用GAPDH做内参的
The doer of good becomes good, the doer of evil becomes evil.
6楼2015-08-27 09:06:32
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祝福。。。
7楼2015-08-27 09:54:34
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岁月微醺

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引用回帖:
6楼: Originally posted by fuyuandj86 at 2015-08-27 09:06:32
是核蛋白浓度低吗?核蛋白浓度低很正常。一般分离以后细胞质蛋白是细胞核蛋白总量的5-10倍。
你用的核蛋白的内参是什么?核蛋白不能用GAPDH做内参的

我用的是lamin B
8楼2015-08-27 12:48:08
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fuyuandj86

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小木虫: 金币+0.5, 给个红包,谢谢回帖
岁月微醺: 金币+10, 谢谢 2015-08-28 10:46:51
引用回帖:
8楼: Originally posted by 岁月微醺 at 2015-08-26 16:48:08
我用的是lamin B...

laminB的确是常见的核蛋白内参,但是分离核蛋白的时候,有的分离方法不容易获得高浓度的laminB、
histone H3 比laminB更常用,一般都能有比较好的信号,只是histone H3的分子量太小了,看大蛋白的时候跑胶不方便
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9楼2015-08-28 05:38:55
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阿Q~~

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路过的看了一下,帮顶,祝福你好运!~~~~~~~~~~~~~~~~~~
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10楼2015-08-28 07:12:47
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