I have used TBP as a loading control in a nuclear extraction of VSMCs, but I noticed it can be changed by NF-KB activity.
Are there any suggestions for other loading control which would not be affected by NF-KB activity.
I think histone proteins. They are present in abundant in nucleus and I feel strongly that their amount won't change drastically to stimulus generally we give to various cell lines over 24 hours. I could not see any change of histone proteins even after 24 hours of curcumin or MPTQ treatment. I hope it will work with you as well. You have to load very less amount of total protein may be only one micro-gram of nuclear extract proteins.
Thank you so much Dr. Ranjit K Giri for your answer and sharing your experience. I will try to check histone and I hope it would work.
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