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Shaking can damage the fragile peptide chains

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The membrane pellet was homogenized and solubilized by gentle nutation for 2 hours at 4 C in buffer consisting of 20 mM Tris-Cl (pH 8.0), 150 mM NaCl, 1% Dodecyl--D-Maltosid (w/v, DDM), and 20 M DNQX, followed by centrifugation at 200,000 g for 30 min to remove insoluble material

PMID: 16987039 | DOI: 10.1089/ars.2006.8.1865 [2] Erden-Inal M, Sunal E, Kanbak G

F., Regulation of Insulin Sensitivity by Serine/Threonine Phosphorylation of Insulin Receptor Substrate Proteins IRS1 and IRS2, Diabetologia 55, no