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A 10 g cell pellet was resuspended in 30 ml buffer B (25 mM Tris-HCl, pH 7.5, 10% glycerol, 0.01% Igepal CA-630, 2 mM 2-Mercaptoethanol) containing 500 mM KCl and a cocktail of protease inhibitors (PIs) (5 g/ml each of aprotinin, chymostatin, leupeptin, and pepstatin A) and lysed by sonication (QSonica Q700) at an amplitude of 40 for two min with 20 s pulses
Using exactly 5 ml of bacteriostatic water with a 5 mg vial gives a clean 1 mg/ml concentration that math makes dose calculation simple and consistent every time you draw a syringe
T., Manghi, F
Lei, G., Zhuang, L
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