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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. and Popp, O. and 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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