I tested two primary antibodies anti-beta-actin so far, and I am getting very inconsistent results accross my Western blots, sometimes I don't even get any detection, and sometimes I get unspecific bands. I am using the same amount of proteins in each well, and I tried to optimize the transfer and incubation time with no success. Any help is highly appreciated.
Other common loading controls include GAPDH and Alpha Tubulin both of which are very strong and specific antibodies in my experience.
Some papers also do a total protein stain as their loading control which could be a ponceau or coomassie depending on if you want to stain at the gel or western blot stage.
I have used antibody against beta-actin from Cell signaling technology and it's great.
Have also used GAPDH from Cell signaling and that worked well as well.
Relatively, beta actin worked better in my case. Both were rabbit Abs.
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