Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation
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. Ye...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2022-05-01
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| Series: | BioTechniques |
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| Online Access: | https://www.future-science.com/doi/10.2144/btn-2021-0074 |
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| author | Gülkiz Baytek Oliver Popp Philipp Mertins Baris Tursun |
| author_facet | Gülkiz Baytek Oliver Popp Philipp Mertins Baris Tursun |
| author_sort | Gülkiz Baytek |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-564122ebeb0549fd830bd5d5627d1925 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 2022-05-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-564122ebeb0549fd830bd5d5627d19252025-08-20T02:25:37ZengTaylor & Francis GroupBioTechniques0736-62051940-98182022-05-0172517518410.2144/btn-2021-0074Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparationGülkiz Baytek0Oliver Popp1Philipp Mertins2Baris Tursun31Berlin Institute for Medical Systems Biology, 10115, Berlin, Germany2Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 10115, Berlin, Germany2Max Delbrück Center for Molecular Medicine in the Helmholtz Association, 10115, Berlin, Germany1Berlin Institute for Medical Systems Biology, 10115, Berlin, GermanyStudying 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.https://www.future-science.com/doi/10.2144/btn-2021-0074C. elegansco-immunoprecipitationFACTHMG-3HMG-4mass spectrometry |
| spellingShingle | Gülkiz Baytek Oliver Popp Philipp Mertins Baris Tursun Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation BioTechniques C. elegans co-immunoprecipitation FACT HMG-3 HMG-4 mass spectrometry |
| title | Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation |
| title_full | Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation |
| title_fullStr | Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation |
| title_full_unstemmed | Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation |
| title_short | Robust co-immunoprecipitation with mass spectrometry for Caenorhabditis elegans using solid-phase enhanced sample preparation |
| title_sort | robust co immunoprecipitation with mass spectrometry for caenorhabditis elegans using solid phase enhanced sample preparation |
| topic | C. elegans co-immunoprecipitation FACT HMG-3 HMG-4 mass spectrometry |
| url | https://www.future-science.com/doi/10.2144/btn-2021-0074 |
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