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1.Cut the tissue in several small pieces, add 10 ml ice chilled lysis buffer (150 mM phosphate buffer pH 7.5, 0.5 % Triton x-100, 1 mM PMSF, 15 ug/ml Aprotinin, 10 ug/ml Leupeptin, 10 ug/ml Pepstatin) and homogenize (best with an electrical tissue homogenizer 3x1min, prevent from warming up through ice-water
bath) until cells are broken.
2.Transfer homogenate into 1.5 ml micro centrifuge tubes and spin for 15 min at 4 deg. Pool supernatant (~ 9 ml) determine protein concentration using a detergent compatible assay.
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