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Inozemtseva LS, et al European Journal of Pharmacology
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The membrane sample containing agonist–MC4R–Gs–Nb35 complex was collected by centrifugation at 46,000× g and carefully resuspended in 20 mM HEPES pH 7.5, 150 mM NaCl, 2 mM MgCl 2 , 1 mM CaCl 2 , 25 µM TCEP, 2.5 mg/mL leupeptin, 0.16 mg/mL bezamidine, 1 µM agonist and 1% n-dodecyl β-d-maltoside (DDM), 0.1% cholesteryl hemisuccinate (CHS) (Anatrace, Inc.)
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