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Solutions: Wash buffer: 50 mM Tris, 5 mM EDTA, 1 g/ml leupeptin, 1 g/ml pepstatin, 0.15 mm PMSF, pH 8.0 U buffer: 5 M urea, 50 mM Tris, 5 mM EDTA,5 mM 2-ME, 1 g/ml leupeptin, 1 g/ml pepstatin, 0.15 mM PMSF, 1 mM DFP pH 8.0 PBS/glycerol: 1X PBS, 20% glycerol, 1% Triton X-100, 5 mM 2-ME, 5 mM EDTA, 5 mM 2- ME, 1 g/ml leupeptin, 1 g/ml pepstatin, 0.15 mM PMSF, 1 mM DFP pH 7.4 21 Use of a peristaltic pump is advised to evenly control flow rates