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JoVE Journal Neuroscience
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Un protocollo migliorato per purificare e direttamente mono-biotinyare recombinante BDNF in un tubo per gli studi sul traffico cellulare nei neuroni

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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Il BDNF ricombinante contenente una sequenza Avi (BDNFAvi) viene prodotto nelle cellule HEK293 in modo conveniente ed è purificato dalla cromatografia di affinità. BDNFavi viene quindi direttamente mono-biotinylated con l'enzima BirA in un tubo. BDNFavi e BDNFavi monobionlate mantengono la loro attività biologica rispetto al BDNF disponibile in commercio.

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Neuroscienze numero 161 BDNF monobio punti quatum traffico assonale purificazione delle proteine dinamica endosomica tracciamento in vivo
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