Protocol to visualize ion channel trafficking in acutely isolated rodent neurons using live-cell immunocytochemistry

STAR Protoc. 2023 Dec 15;4(4):102744. doi: 10.1016/j.xpro.2023.102744. Epub 2023 Nov 23.

Abstract

Here, we present a protocol for live-cell immunocytochemistry to demonstrate reversible translocation of ion channels to the neuronal cell surface. We describe steps for cell preparation and isolation, experimental treatment, antibody binding prior to fixation, specific pipetting techniques, troubleshooting, and expected outcomes of correct use of the protocol. This protocol will be useful to study regulated translocation of ion channels and other membrane proteins. For complete details on the use and execution of this protocol, please refer to Haan et al.1.

Keywords: Cell culture; Cell isolation; Microscopy; Neuroscience.

MeSH terms

  • Cell Membrane
  • Immunohistochemistry
  • Ion Channels*
  • Membrane Proteins*
  • Neurons

Substances

  • Ion Channels
  • Membrane Proteins