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Western blot The total protein extracts were prepared using RIPA buffer (50 mM Tris, 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), 1 mM phenylmethylsulfonyl fluoride (PMSF), 1 mM Na3VO4, 5 mM NaF, and 1% cocktail protein protease inhibitors (Sigma)
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Hydrogen atoms were added using Chimera 1.14 32
Similarly, a preparation of calcitonin receptor (CTR)-calcitonin-Gs complex did not resolve clear density for the ECD of CTR attributed to partial flexibility 8
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