Z******5 发帖数: 435 | 1 要检测的蛋白质是个转录因子,细胞核内外都有。
用全细胞蛋白去做WB,信号很弱,想用细胞核蛋白做一下试试。 |
c**k 发帖数: 1228 | 2 use lamin-a or HDAC1
【在 Z******5 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 要检测的蛋白质是个转录因子,细胞核内外都有。 : 用全细胞蛋白去做WB,信号很弱,想用细胞核蛋白做一下试试。
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k*****n 发帖数: 323 | |
T****u 发帖数: 424 | |
Z******5 发帖数: 435 | 5 因为目前手头只有actin的抗体,想问问用这个行不行。 |
k*****n 发帖数: 323 | |
A******y 发帖数: 2041 | 7 If you are doing nuclear extract and you can see actin...then your technique
is not perfect yet. |
b**j 发帖数: 415 | 8 maybe you can, if no actin signal for nuclear fraction, then your extraction
is good |
W****C 发帖数: 1937 | 9
technique
这从何说起。。。核里的actin也不少。。。。
他是要做内参, 不是要做marker, 如果真是不舍得买histone的抗体,只是做个预
实验的话我觉得可以, 但是要发paper的话可能不被接受--抗体都不舍得买,
editor肯定不买账。。。。
【在 A******y 的大作中提到】![](/moin_static193/solenoid/img/up.png) : If you are doing nuclear extract and you can see actin...then your technique : is not perfect yet.
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A******y 发帖数: 2041 | 10 There is actin in the nucleus? I did a lot cyto and nuclear extracts and using sigma actin ab (the strongest one out there) you can barely see a band while the cyto is like 1 sec exposure...Submit your paper with actin as nuclear loading control, let's see what the
review will say.
【在 W****C 的大作中提到】![](/moin_static193/solenoid/img/up.png) : : technique : 这从何说起。。。核里的actin也不少。。。。 : 他是要做内参, 不是要做marker, 如果真是不舍得买histone的抗体,只是做个预 : 实验的话我觉得可以, 但是要发paper的话可能不被接受--抗体都不舍得买, : editor肯定不买账。。。。
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s******y 发帖数: 28562 | 11 核里到底有没有actin 一直是领域里面在争吵的问题,
好像是有ARP (actin-related protein),然后据说在某些特定情况下
actin 会进去。
当然,肯定不能用用这个作为loading control.
using sigma actin ab (the strongest one out there) you can barely see a
band while the cyto is like 1 sec exposure...Submit your paper with actin as
nuclear loading control, let's see what the
【在 A******y 的大作中提到】![](/moin_static193/solenoid/img/up.png) : There is actin in the nucleus? I did a lot cyto and nuclear extracts and using sigma actin ab (the strongest one out there) you can barely see a band while the cyto is like 1 sec exposure...Submit your paper with actin as nuclear loading control, let's see what the : review will say.
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s*****g 发帖数: 7857 | 12 检查出actin多了只能说你的核蛋白污染了细胞质蛋白.通常要用组蛋白等做LAODING
CONTROL, 再用actin/GAPDH等做污染检测.
【在 Z******5 的大作中提到】![](/moin_static193/solenoid/img/up.png) : 因为目前手头只有actin的抗体,想问问用这个行不行。
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