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Optimized protocol for in vivo affinity purification proteomics and biochemistry using C. elegans

Desbois, Muriel; Pak, Joseph S.; Opperman, Karla J.; Giles, Andrew C.; Grill, Brock

Authors

Joseph S. Pak

Karla J. Opperman

Andrew C. Giles

Brock Grill



Abstract

We present an optimized protocol for in vivo affinity purification proteomics and biochemistry using the model organism C. elegans. We describe steps for target tagging, large-scale culture, affinity purification using a cryomill, mass spectrometry and validation of candidate binding proteins. Our approach has proven successful for identifying protein-protein interactions and signaling networks with verified functional relevance. Our protocol is also suitable for biochemical evaluation of protein-protein interactions in vivo.

For complete details on the use and execution of this protocol, please refer to Crawley et al.,1 Giles et al.,2 and Desbois et al.3

Journal Article Type Article
Acceptance Date Jun 8, 2023
Online Publication Date Jun 8, 2023
Publication Date Jun 16, 2023
Deposit Date Nov 20, 2023
Journal STAR Protocols
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 4
Issue 2
DOI https://doi.org/10.1016/j.xpro.2023.102262
Publisher URL https://www.sciencedirect.com/science/article/pii/S2666166723002204?via%3Dihub


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