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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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小木虫: 金币+0.5, 给个红包,谢谢回帖
给的信息太少,不好分析。
2楼2015-08-25 17:15:23
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gJordys

新虫 (初入文坛)

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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+5 2015-08-27 12:46:14
Antitumor and immunomodulating activity of a polysaccharide from Sophora flavescens Ait.
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International Journal of Biological Macromolecules, Volume 51, Issue 5, December 2012, Pages 705-709
Lu Bai, Li-Ying Zhu, Bao-Shan Yang, Li-Jun Shi, Yong Liu, Ai-Min Jiang, Li-Li Zhao, Guang Song, Tie-Fu Liu
Immunological response in H22 transplanted mice undergoing Aconitum coreanum polysaccharide treatment
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International Journal of Biological Macromolecules, Volume 55, April 2013, Pages 295-300
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Antitumor and antiangiogenic activity of Schisandra chinensis polysaccharide in a renal cell carcinoma model
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Immunopontentiating and antitumor activities of a polysaccharide from Pulsatilla chinensis (Bunge) Regel
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Tong Liu, Leiguang Ye, Xueqing Guan, Xiaoshuan Liang, Cong Li, Qian Sun, Yang Liu, Shiru Chen, Fa Bang, Baogang Liu
Panax ginseng polysaccharide suppresses metastasis via modulating Twist expression in gastric cancer
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Jian-Ping Cai, Yang-Jiazi Wu, Cong Li, Mei-Yan Feng, Qing-Tao Shi, Rui Li, Zhen-Yu Wang, Jing-Shu Geng
Purification, characterization and anticancer activity of a polysaccharide from Panax ginseng
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International Journal of Biological Macromolecules, Volume 51, Issue 5, December 2012, Pages 968-973
Cong Li, Jianping Cai, Jingshu Geng, Yinghong Li, Zhenyu Wang, Rui Li
3楼2015-08-25 19:42:29
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gJordys

新虫 (初入文坛)

Synergetic effect of Andrographis paniculata polysaccharide on diabetic nephropathy with andrographolide
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International Journal of Biological Macromolecules, Volume 51, Issue 5, December 2012, Pages 738-742
Jie Xu, Zhanting Li, Min Cao, Han Zhang, Jifeng Sun, Jie Zhao, Qiang Zhou, Zhifen Wu, Lianjia Yang
Anti-hepatoma activity and mechanism of corn silk polysaccharides in H22 tumor-bearing mice
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Jingyue Yang, Xiao Li, Yan Xue, Nan Wang, Wenchao Liu
A polysaccharide from Sanguisorbae radix induces caspase-dependent apoptosis in human leukemia HL-60 cells
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Zhigang Wu, Honghui Sun, Jingzhong Li, Chijiao Ma, Siqiao Zhao, Zheng Guo, Yao Lin, Yaping Lin, Li Liu
Immunopontentiating activities of the purified polysaccharide from evening primrose in H22 tumor-bearing mice
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International Journal of Biological Macromolecules, Volume 52, January 2013, Pages 280-285
Guang Zeng, Ying Ju, Huan Shen, Ningxin Zhou, Li Huang
Auricularia polytricha polysaccharides induce cell cycle arrest and apoptosis in human lung cancer A549 cells
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Jie Yu, Ruilin Sun, Zhongquan Zhao, Yingyu Wang
4楼2015-08-25 19:43:32
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gJordys

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小木虫: 金币+0.5, 给个红包,谢谢回帖
Antioxidant activities of different polysaccharide conjugates (CRPs) isolated from the fruiting bodies of Chroogomphis rutilus (Schaeff.: Fr.) O. K. MillerOriginal Research Article
Carbohydrate Polymers, Volume 82, Issue 2, 5 September 2010, Pages 510-514
Yong-Xu Sun, John F. Kennedy

Purification, structural analysis and hydroxyl radical-scavenging capacity of a polysaccharide from the fruiting bodies of Russula virescensOriginal Research Article
Process Biochemistry, Volume 45, Issue 6, June 2010, Pages 874-879
Yong-Xu Sun, Ji-Cheng Liu, Xiu-Dong Yang, John F. Kennedy
5楼2015-08-25 19:45:32
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fuyuandj86

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小木虫: 金币+0.5, 给个红包,谢谢回帖
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

金虫 (小有名气)

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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己所不欲,勿施于人

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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
The doer of good becomes good, the doer of evil becomes evil.
9楼2015-08-28 05:38:55
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阿Q~~

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小木虫: 金币+0.5, 给个红包,谢谢回帖
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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