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The cell pellet was resuspended and lysed in lysis buffer (20 mM HEPES-NaOH pH 7.4, 200 mM NaCl, 1% Triton-X100 (v/v), 2 mM MgCl 2 , 0.5 mM TCEP) with protease inhibitors (1 mM PMSF, 1 M pepstatin, 4 M leupeptin, 0.3 M aprotinin) for 30 min at 4 C
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The delay in implementation is to provide an additional year for broad communication and education of athletes, their entourage and medical personnel so that there is a better understanding of the practical implementation of tramadol prohibition in competition
7 BernardG.ShevellM.I (2008)
