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Protein Purification-freie Methode der Bindungsaffinität Bestimmung durch Microscale Thermophorese
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Protein Purification-freie Methode der Bindungsaffinität Bestimmung durch Microscale Thermophorese

Article DOI: 10.3791/50541-v • 10:22 min • August 15th, 2013
August 15th, 2013 •
1,2, 1, 3, 4, 1, 1, 1
1Cancer and Inflammation Program, National Cancer Institute, 2Basic Science Program, SAIC-Frederick, Inc., 3Departments of Oncology and Radiation Medicine, Georgetown University Medical Center, 4Structural Biophysics Laboratory, National Cancer Institute

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Microscale Thermophorese (MST) kann in weiten Bereichen zur Bestimmung der Bindungsaffinität werden ohne Reinigung des Zielproteins aus Zelllysaten. Das Protokoll umfasst die Überexpression des GFP-Fusionsprotein, Zell-Lyse in nicht-denaturierenden Bedingungen, sowie Erkennung MST Signal in der Gegenwart von variierenden Konzentrationen des Liganden.

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Molecular Biology Ausgabe 78 Biochemie Zellbiologie Genetik Chemie Pharmakologie intrazelluläre Signaltransduktion Peptide und Proteine Proteine Protein-Inhibitor-Wechselwirkung K Transkriptionsfaktor Ligandenbindung Bindungsaffinität Thermophorese Fluoreszenz-Mikroskopie
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