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A toolbox of nanobodies developed and validated for use as intrabodies and nanoscale immunolabels in mammalian brain neurons.
Dong, Jie-Xian; Lee, Yongam; Kirmiz, Michael; Palacio, Stephanie; Dumitras, Camelia; Moreno, Claudia M; Sando, Richard; Santana, L Fernando; Südhof, Thomas C; Gong, Belvin; Murray, Karl D; Trimmer, James S.
Afiliación
  • Dong JX; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Lee Y; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Kirmiz M; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Palacio S; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Dumitras C; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Moreno CM; Department of Physiology and Membrane Biology, University of California, Davis, Davis, United States.
  • Sando R; Department of Physiology and Biophysics, University of Washington, Seattle, United States.
  • Santana LF; Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford School of Medicine, Stanford, United States.
  • Südhof TC; Department of Physiology and Membrane Biology, University of California, Davis, Davis, United States.
  • Gong B; Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford School of Medicine, Stanford, United States.
  • Murray KD; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
  • Trimmer JS; Department of Neurobiology, Physiology and Behavior, University of California, Davis, Davis, United States.
Elife ; 82019 09 30.
Article en En | MEDLINE | ID: mdl-31566565
Nanobodies (nAbs) are small, minimal antibodies that have distinct attributes that make them uniquely suited for certain biomedical research, diagnostic and therapeutic applications. Prominent uses include as intracellular antibodies or intrabodies to bind and deliver cargo to specific proteins and/or subcellular sites within cells, and as nanoscale immunolabels for enhanced tissue penetration and improved spatial imaging resolution. Here, we report the generation and validation of nAbs against a set of proteins prominently expressed at specific subcellular sites in mammalian brain neurons. We describe a novel hierarchical validation pipeline to systematically evaluate nAbs isolated by phage display for effective and specific use as intrabodies and immunolabels in mammalian cells including brain neurons. These nAbs form part of a robust toolbox for targeting proteins with distinct and highly spatially-restricted subcellular localization in mammalian brain neurons, allowing for visualization and/or modulation of structure and function at those sites.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Coloración y Etiquetado / Encéfalo / Transporte de Proteínas / Anticuerpos de Dominio Único / Neuronas Límite: Animals Idioma: En Revista: Elife Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Coloración y Etiquetado / Encéfalo / Transporte de Proteínas / Anticuerpos de Dominio Único / Neuronas Límite: Animals Idioma: En Revista: Elife Año: 2019 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Reino Unido