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The Rho-kinase activity was evaluated by measuring its substrate phosphorylation (pMYPT1) using 100 µg of muscle lysate, 1 mM of ATP, and 500 ng of recombinant MYPT1 (Millipore®). This mixture was incubated for 30 min under agitation at 30°C. The reaction was stopped using 2x Laemmli and submitted to IP protocol. The membrane was incubated using anti-pMYPT1 (T696) from Millipore®.
Copyright and License information: ©2021 Muñoz, Gaspar, Severino, Macêdo, Simabuco, Ropelle, Cintra, da Silva, Kim and Pauli
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