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Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation

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Item Type:Article
Title:Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation
Creators Name:Baytek, G., Popp, O., Mertins, P. and Tursun, B.
Abstract:Studying protein interactions in vivo can reveal key molecular mechanisms of biological processes. Co-immunoprecipitation with mass spectrometry detects protein–protein interactions with high throughput. The nematode Caenorhabditis elegans is a powerful genetic model organism for in vivo studies. Yet its rigid and complex tissues require optimization for biochemistry applications to ensure reproducibility. The authors optimized co-immunoprecipitation with mass spectrometry by combining a native co-immunoprecipitation procedure with single-pot, solid-phase enhanced sample preparation. The authors' results for the highly conserved chromatin regulator FACT subunits HMG-3 and HMG-4 demonstrated that single-pot, solid-phase enhanced sample preparation-integrated co-immunoprecipitation with mass spectrometry procedures for C. elegans samples are highly robust. Moreover, in an accompanying study about the chromodomain factor MRG-1 (MRG15 in humans), the authors demonstrated remarkably high reproducibility for ten replicate experiments.
Keywords:C. elegans, Co-Immunoprecipitation, FACT, HMG-3, HMG-4, Mass Spectrometry, Protein-Protein Interaction, SP3, Animals, Caenorhabditis elegans
Source:Biotechniques
ISSN:0736-6205
Publisher:Future Science
Volume:72
Number:5
Page Range:175-184
Date:May 2022
Official Publication:https://doi.org/10.2144/btn-2021-0074
PubMed:View item in PubMed

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