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JoVE Journal Neuroscience
En forbedret protokoll for å rense og direkte mono-bio-biotinylate rekombinant BDNF i et rør for cellulære trafficking studier i nevroner
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En forbedret protokoll for å rense og direkte mono-bio-biotinylate rekombinant BDNF i et rør for cellulære trafficking studier i nevroner

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 som inneholder en Avi-sekvens (BDNFAvi) produseres i HEK293-celler på en kostnadseffektiv måte og renses av affinitetkromatografi. BDNFavi blir deretter direkte mono-biotinylert med enzymet BirA i et rør. BDNFavi og mono-biotinylated BDNFavi beholder sin biologiske aktivitet sammenlignet med kommersielt tilgjengelig BDNF.

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Nevrovitenskap Utgave 161 BDNF mono-biotinylation quatum prikker axonal smugling protein rensing endosome dynamikk in vivo sporing
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