I have some cells that I know are proliferating less after drug treatment however are not showing a decrease in PCNA by Western Blot, I have read in a number of places now that PCNA can also be expressed by resting cells and may not be the best option in my case. Ki67 is not really an option due to it's large size but what do you use as a proliferation marker by WB? Thanks in advance!
One can look at a number of proteins by western blotting- including cyclins D1, A, and E levels, and E2F1.
Hi Rachel,
Please tell me the detail protocol what you have done for this expt?
which cells used for this experiment?
what is the objective of your experiment?
Although you got the marker transferred to the membrane, I suggest you first check for the presence of proteins with an antibody which should give you a signal (like a-Tubulin, GAPDH, Actin, ...).
If you got your proteins transferred to the membrane, you can try to increase the exposure time as your target might have a very low concentration.
Hope this helps
Perfect thank you everyone! I have some of those antibodies so will give Cyclin D1 and p21 a go.
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