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

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

Citation

Desbois, M., Pak, J. S., Opperman, K. J., Giles, A. C., & Grill, B. (2023). Optimized protocol for in vivo affinity purification proteomics and biochemistry using C. elegans. STAR Protocols, 4(2), https://doi.org/10.1016/j.xpro.2023.102262

• 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