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After washing the plates five times with TBS + 0.05% Tween-20, plates were developed by adding develop solution (1% 3,3,5,5-tetramethylbenzidine (TMB, Sigma-Aldrich), 0.01 % H 2 O 2 , 100 mM sodium acetate, 100 mM citric acid) and the reaction was stopped after 3 min by adding 0.8 M H 2 SO 4

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ISSN 0002-9394

The identity of these metabolites was confirmed by tandem mass spectrometry

ACE-031 is a myostatin inhibitor designed to promote muscle growth by blocking myostatin, a protein that limits muscle development