Site-specific direct labeling of neurotrophins and their receptors: From biochemistry to advanced imaging applications

Authors:

Gobbo, F., Bonsignore, F., Amodeo, R., Cattaneo, A., & Marchetti, L.

Keywords:

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

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.

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