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

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.

SUBMITTER: Desbois M 

PROVIDER: S-EPMC10323129 | biostudies-literature | 2023 Jun

REPOSITORIES: biostudies-literature