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Western immunoblot analyses For the preparation of total liver homogenates, frozen liver pieces were homogenized in RIPA buffer (150 mM NaCl, 1% TritonX-100, 0.25% sodium deoxycholate, 0.1% SDS, 50 mM Tris, 1 mM EDTA, 1 mM PMSF, protease inhibitor cocktail, pH 7.4) on ice with a tissue tearor (BioSpec Products, Bartlesville)
10.1007/s00424-015-1768-3 Pflugers
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At varying time points (0, 2, 5, 15, 30 min), purified Pt GSTF8 (8 M) and GSH (20 M) were added to an aliquot of the equilibration reaction and the resulting mixtures were incubated at 30 C for 30 min at 1200 rpm using a 3 mm shaking diameter before extraction with 2 vol of 75% methanol in H 2 O, 1 mM EDTA, 0.3% hydrochloric acid
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