Hey all,

I struggle a lot with Blue Native. The protein I want to look at is localized at the PM and forms large oligomers. However, I just can't extract it. I've tried it with:

Buffer 1:

10% glycerol, 50 mM HEPES (pH 7.4), 5 mM MgCl2, 50 mM NaCl, 10 mM DTT, 1x protease inhibitor cocktail, 1% Digitonin

and

Buffer 2:

50 mM Tris-Cl, pH 7.5, 50 mM NaCl, 5 mM MgCl2, 10% (v/v) Glyceroll, 10 mM DTT

0.2% (v/v) NP-40, Protease inhibitor cocktail

Any other ideas / protocol that worked for PM proteins?

I would really appreciate it :)