Protocol for the purification and analysis of nuclear UFMylated proteins
Summary: Protein UFMylation regulates numerous cellular processes including ribosome quality control and nuclear DNA repair. Here, we present a technique to isolate nuclei and purify UFMylated proteins under denaturing non-reducing conditions from commonly used mammalian cell line models such as hTE...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725000401 |
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| author | Pudchalaluck Panichnantakul Marlene Oeffinger |
| author_facet | Pudchalaluck Panichnantakul Marlene Oeffinger |
| author_sort | Pudchalaluck Panichnantakul |
| collection | DOAJ |
| description | Summary: Protein UFMylation regulates numerous cellular processes including ribosome quality control and nuclear DNA repair. Here, we present a technique to isolate nuclei and purify UFMylated proteins under denaturing non-reducing conditions from commonly used mammalian cell line models such as hTERT-RPE1, HEK293, U2OS, and HCT116 cells. We then describe procedures for identifying and analyzing purified UFMylated proteins using mass spectrometry and western blot.For complete details on the use and execution of this protocol, please refer to Panichnantakul et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-563e7cbffd8e4f68919681a11932a106 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-563e7cbffd8e4f68919681a11932a1062025-02-12T05:32:49ZengElsevierSTAR Protocols2666-16672025-03-0161103634Protocol for the purification and analysis of nuclear UFMylated proteinsPudchalaluck Panichnantakul0Marlene Oeffinger1Institut de Recherches Cliniques de Montréal, Center for Genetic and Neurological Diseases, 110 Avenue des Pins Ouest, Montréal, Québec H2W 1R7, Canada; Division of Experimental Medicine, Faculty of Medicine, McGill University, Montréal, Québec H4A 3J1, CanadaInstitut de Recherches Cliniques de Montréal, Center for Genetic and Neurological Diseases, 110 Avenue des Pins Ouest, Montréal, Québec H2W 1R7, Canada; Division of Experimental Medicine, Faculty of Medicine, McGill University, Montréal, Québec H4A 3J1, Canada; Département de Biochimie et Médicine Moléculaire, Faculté de Médicine, Université de Montréal, Montréal, Québec H3C 3J7, Canada; Corresponding authorSummary: Protein UFMylation regulates numerous cellular processes including ribosome quality control and nuclear DNA repair. Here, we present a technique to isolate nuclei and purify UFMylated proteins under denaturing non-reducing conditions from commonly used mammalian cell line models such as hTERT-RPE1, HEK293, U2OS, and HCT116 cells. We then describe procedures for identifying and analyzing purified UFMylated proteins using mass spectrometry and western blot.For complete details on the use and execution of this protocol, please refer to Panichnantakul et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725000401Cell BiologyCell separation/fractionationProteomics |
| spellingShingle | Pudchalaluck Panichnantakul Marlene Oeffinger Protocol for the purification and analysis of nuclear UFMylated proteins STAR Protocols Cell Biology Cell separation/fractionation Proteomics |
| title | Protocol for the purification and analysis of nuclear UFMylated proteins |
| title_full | Protocol for the purification and analysis of nuclear UFMylated proteins |
| title_fullStr | Protocol for the purification and analysis of nuclear UFMylated proteins |
| title_full_unstemmed | Protocol for the purification and analysis of nuclear UFMylated proteins |
| title_short | Protocol for the purification and analysis of nuclear UFMylated proteins |
| title_sort | protocol for the purification and analysis of nuclear ufmylated proteins |
| topic | Cell Biology Cell separation/fractionation Proteomics |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725000401 |
| work_keys_str_mv | AT pudchalaluckpanichnantakul protocolforthepurificationandanalysisofnuclearufmylatedproteins AT marleneoeffinger protocolforthepurificationandanalysisofnuclearufmylatedproteins |