What's the reasonable loading control of nuclear protein in acute injury? Is HDAC1 or Histone H3 a resonable loading control of nuclear protein in acute injury model? How to explain total my total protein of a specific signaling pathway is almost the same, but HDAC1 or Histone H3 various from each other? I have performed BCA assay for nuclear protein concentration and adjust them to the same loading volume (20 ug) in my Western blot.
Looking forward to your suggestions.
I have the same problem. Nether H3 or HDAC1 are housekeeping proteins in my case. I used cell number at the start of sample preparation and DNA amount after sample is prepared but the latter is also no good for longer then 8h treatment because cells can stop replicating if injured. Do experiment as a triplicate and blot for different proteins. When you find ones that do not change use them as a loading control (try H3, actin, lamins, PCNA...)
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