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JoVE Journal Neuroscience
En forbedret protokol til at rense og direkte Mono-Biotinylate Recombinant BDNF i et rør for Cellular Trafficking Studies i Neuroner
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En forbedret protokol til at rense og direkte Mono-Biotinylate Recombinant BDNF i et rør for Cellular Trafficking Studies i Neuroner

Article DOI: 10.3791/61262-v • 13:46 min • July 11th, 2020
July 11th, 2020 •
1,2, 1,3, 1,3, 4, 1,3
1Department of Physiology, Faculty of Biological Sciences, Pontificia Universidad Católica de Chile, 2Department of Molecular and Cellular Biology, Faculty of Biological Sciences, Center for Aging and Regeneration (CARE UC), Pontificia Universidad Católica de Chile, 3Institute of Biomedical Sciences. Faculty of Medicine and Faculty of Life Sciences, Universidad Andrés Bello, 4Department of Neuromuscular Diseases, UCL Queen Square Institute of Neurology and UK Dementia Research Institute at UCL, University College London Campus

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Rekombinant BDNF indeholdende en Avi-sekvens (BDNFAvi) fremstilles i HEK293-celler på en omkostningseffektiv måde og renses ved affinitetskromatografi. BDNFavi monobiotinyliseres derefter direkte med enzymet BirA i et rør. BDNFavi og mono-biotinyleret BDNFavi bevarer deres biologiske aktivitet sammenlignet med kommercielt tilgængelige BDNF.

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Neurovidenskab BDNF mono-biotinylering quatum prikker axonal handel protein rensning endelsesdynamik in vivo tracking
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