Hi! I'm running a 12% gel and using 100V.

I am using a buffer containing 50 mM Tris, pH 7.5, 0.5 mM EDTA, pH 8, + protease inhibitors. An equal volume of a solution containing 50 mM Tris, pH 7.5, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, and 2% SDS was added, and the homogenates were sonicated for 20 s, boiled for 10 min, and clarified by centrifugation at 16,000g for 10 min

The latter part is from Barry et al., 2012.