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Eiwitzuivering-vrij bindwijze affiniteit Bepaling door Microschaal thermophoresis

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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Microschaal thermophoresis (MST) op grote schaal kunnen worden gebruikt voor de bepaling van de bindingsaffiniteit zonder zuivering van het doelwit eiwit uit cellysaten. Het protocol omvat overexpressie van GFP-gefuseerde eiwit, cellyse in niet-denaturerende omstandigheden en detectie van MST signaal in de aanwezigheid van variërende concentraties van het ligand.

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Molecular Biology biochemie celbiologie genetica chemie farmacologie intracellulaire signalering peptiden en proteïnen eiwitten eiwit-remmer interactie K Transcriptiefactor ligand-binding bindingsaffiniteit thermophoresis fluorescentie microscopie
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