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Site-Specific Direct Labeling of Neurotrophins and Their Receptors: From Biochemistry to Advanced Imaging Applications

  • Francesco Gobbo
  • Fulvio Bonsignore
  • Rosy Amodeo
  • Antonino Cattaneo
  • Laura Marchetti
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1727)

Abstract

We describe here a versatile methodological platform to achieve site-directed and stoichiometry-controlled labeling of neurotrophins and their receptors with various probes, ranging from biotin to small organic dyes. This labeling method works in vitro on purified neurotrophins as well as in a living cell context, where it achieves selective labeling of surface-exposed neurotrophin receptors. Here, we list all experimental details of our labeling protocols, along with examples of the wide range of applications in which these can be used.

Key words

Neurotrophin Nerve growth factor TrkA tyrosine kinase Membrane receptor pool Fluorescence labeling Biotinylation Axonal transport Inducible lentivirus Neuron culture Astrocyte culture 

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Francesco Gobbo
    • 1
  • Fulvio Bonsignore
    • 2
  • Rosy Amodeo
    • 2
    • 3
  • Antonino Cattaneo
    • 1
  • Laura Marchetti
    • 1
    • 2
    • 3
  1. 1.Bio@SNS, Scuola Normale SuperiorePisaItaly
  2. 2.NEST, Scuola Normale SuperiorePisaItaly
  3. 3.Center for Nanotechnology Innovation @NEST, Istituto Italiano di TecnologiaPisaItaly

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