Hi everyone,
I am treating differentiated C2C12s with Doxorubicin. I probed for phospho and total Akt. Unlike T-Erk and T-Stat which remained fairly constant, the T-Akt is changing and I would like to know why this is happening? It makes it difficult to normalize the phospho to the total as the values I obtain do not correspond to the image of my bands. Is T-Akt supposed to change and why is this happening? Is it ok, in this scenario, to only present the phospho and only the total, instead of the ratio? What is the correct way for analyzing/presenting phosphos and totals? Please please help!
How long is your treatment? If it is long-term, maybe you really have a modification of AKT expression levels. Try actin or tubulin to see if you have variation. If not, you can use this to show your variation in phospho and total AKT.
Hi Roberto
Thank you for the reply and I am sorry I have taken so long to get back to you.
My treatment is every day, for 5 days, however, I see the first increase in total protein after 24 hours. While my GAPDH is fairly constant, it is not perfect, but even after normalizing I see the same pattern. Do you have any articles that mention a similar thing?
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