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Visualizing Wnt Palmitoylation in Single Cells

  • Xinxin Gao
  • Rami N. HannoushEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1481)

Abstract

Wnt palmitoylation regulates its secretion and signaling activity in cells. Methods to monitor cellular Wnt palmitoylation are instrumental in investigating Wnt activity, secretion, and its interaction with cellular membrane compartments. This protocol describes a method we have recently developed to detect cellular Wnt palmitoylation. The method, combining click chemistry, bio-orthogonal fatty acid probes, and proximity ligation assay (PLA), provides high sensitivity and subcellular resolution for detection of Wnt palmitoylation. It is also compatible with multiple imaging platforms, and is applicable to detecting palmitoylated forms of other fatty acylated proteins.

Key words

Wnt palmitoylation Alkyne fatty acid Click chemistry Proximity ligation Microscopy 

Notes

Acknowledgment

We acknowledge use of microscopes at the Center for Advanced Light Microscopy (CALM) at Genentech.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Early Discovery BiochemistryGenentechSouth San FranciscoUSA

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