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Is beta-actin a good cyto vs. nuc control?


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#1 tfcheng

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Posted 26 January 2010 - 06:51 PM

Hi,

Recently I did a bunch of cyto vs. nuclear fractionation, and I used beta-actin to see how my prep is, I got a blot where actin is only detected in cytoplasm, and nearly undetectable in nuclei. But then I read about some articles that seems to me that beta-actin is also present in the nucleus, I am a little confused....


TFC

#2 Radish

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Posted 28 January 2010 - 05:20 PM

Hi,

Recently I did a bunch of cyto vs. nuclear fractionation, and I used beta-actin to see how my prep is, I got a blot where actin is only detected in cytoplasm, and nearly undetectable in nuclei. But then I read about some articles that seems to me that beta-actin is also present in the nucleus, I am a little confused....


TFC


Hello
I have used beta-actin as a loading control for nuclear extracts quite a few times and never had problems.
Are you sure that you have enough protein in the extract to elicit antibody binding?


good luck

#3 bob1

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Posted 31 January 2010 - 03:38 PM

Actin is found in both cytoplasm and nucleus. As a fractionation control try nucleolin or histone.




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