Normally in my laboratory, I use an antibody which detects Total Erk (both: the phosphorylated and the non-phosphorylated kinase) as a loading control for western blot, in apoptosis studies.
Can I use this protein as a load control?
This argument is valid when there is a comparable number of viable cells at all time points. How can I support the use of Erk as a loading control (western Blot) in apoptosis studies?
I have seen ERK2 being used as loading control in certain publications. Theres no evidence ( as far as I know) that ERK1/2 levels are altered during apoptosis. So it should be acceptable.
In certain cases it makes more sense to use something like this, compared to highly expressed actin / tubulin which any way will not change if even 10% cells disappear. If u want to be sure u can do a onetime comparison between erk levels to that of a std loading control (gapdh/actin/tubulin) with the extreme stimulations which u can have.
Good Luck
For loading purposes any housekeeping protein would do. ERKs are quite robust for this purpose but perhaps actin or tubulin are better as they show little fluctuation
Is ERK better than beta actin as western blottting loading control in adipose tissue samples of obese patients?
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