Housekeeping proteins for western blot that do not change following nerve injury

Experimental Techniques
1

Hi all,

I need to run a western blot on protein lysate extracted from mouse DRGs. One of my groups has undergone a spared nerve injury and I’m aware that certain commonly used housekeeping proteins can be up or downregulated following injury.

Can anyone recommend me a housekeeping protein that does not change following nerve injury? I am looking 3-weeks post SNI.

I have checked the Renthanl sequencing data set to get some idea of which proteins might be altered follwing nerve injury. Its seems Beta-Tubulin and Beta-Actin would not be a good option because they are upregulated but GAPDH should be ok for normalisation at 3 weeks.

Thanks in advance for your help.

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Maybe Vinculin?
Much better would be total protein normalization, and then you neednt worry about housekeeping genes changing. https://www.youtube.com/watch?v=N97BQXz4-dY
This is what my lab does. Good luck!

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I do western blots in similar samples and have had luck with GAPDH and total H3 antibodies. Hope this helps.

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Late to the party, but I just wanted to add that Beta3 tubulin and Actin always worked well in my hands. In my experience, most of the variation more often comes from the dissection, meaning how much nerve is left attached to the DRG. We identify the part of the DRG containing the neuronal bodies (it is kind of transparent) and we quickly trim around that. Keeping this consistent is also key for getting good, healthy cultures.