Surface biotinylation of primary neurons to monitor changes in AMPA receptor surface expression in response to kainate receptor stimulation

对原代神经元进行表面生物素化,以监测 AMPA 受体表面表达在海人酸受体刺激后的变化

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作者:Jithin D Nair, Jeremy M Henley, Kevin A Wilkinson

Abstract

Here, we detail a surface biotinylation technique used to label surface-expressed proteins in primary neuronal cultures. Surface proteins are labeled with membrane-impermeant Sulfo-NHS-SS-biotin, and isolated by pull-down with streptavidin beads followed by western blotting to measure levels of surface expression of the protein of interest under different conditions. We have used this approach extensively to monitor activity-dependent changes in α-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and kainate receptor (KAR) subunits. However, this protocol can be used to investigate any surface-expressed protein. For complete details on the use and execution of this protocol, please refer to Nair et al. (2021).

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