A subscription to JoVE is required to view this content. Sign in or start your free trial.
5,607 Views
•
13:46 min
•
July 11, 2020
DOI:
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
Summary
Automatically generated
Avi配列を含む組換えBDNF(BDNFAvi)は、コスト効率の高い方法でHEK293細胞で産生され、アフィニティークロマトグラフィーによって精製される。BDNFaviは、その後、チューブ内の酵素BirAで直接モノビオチン化されます。BDNFaviおよびモノビオチン化BDNFaviは、市販のBDNFと比較して生物学的活性を保持します。
Read Article
Cite this Article
Stuardo, N., Moya-Alvarado, G., Ramírez, C., Schiavo, G., Bronfman, F. C. An Improved Protocol to Purify and Directly Mono-Biotinylate Recombinant BDNF in a Tube for Cellular Trafficking Studies in Neurons. J. Vis. Exp. (161), e61262, doi:10.3791/61262 (2020).
Copy