Hi, I'm trying to measure the relative abundances of complex II of the ETC with a western blot. I know PVDF is best for low abundance proteins and nitrocellulose is best for low background noise, but I don't know what I should prioritize here. Any help would be greatly appreciated.
Overnight wet-blotting using PVDF membrane activated in methanol or ethanol is normally suggested for membrane-anchored or transmembrane containing proteins. For soluble proteins (in 30-150 kDa size range), semi-dry blotting with nitrocellulose membrane works best on my hand.
Good luck!
I would go with nitrocellulose and wet blotting first. The nitrocellulose membrane is much cleaner with the majority of 1 and 2 antibodies. Mostly, I have gotten really nice bands with different protein sizes (11-250kD) and compartments using the nitrocellulose membrane. Good luck
I would propose PVDF membranes. Moreover, Western blotting is NO quantitative method! You may quantify your results afterward. However, this will always be semiquantitative - and unnecessary at all. When expecting quantitative Data, you should perform more adequate methods (e.g. ELISA).
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