Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages
Summary: Protein synthesis, or mRNA translation, is the biological process through which genetic information stored in messenger RNAs is encoded into proteins. Here, we present an optimized protocol for assessing the translation rate in mouse adult microglia and cultured bone-marrow-derived macropha...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723005269 |
| _version_ | 1797685244300623872 |
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| author | Ignazio Antignano Lily Keane Melania Capasso |
| author_facet | Ignazio Antignano Lily Keane Melania Capasso |
| author_sort | Ignazio Antignano |
| collection | DOAJ |
| description | Summary: Protein synthesis, or mRNA translation, is the biological process through which genetic information stored in messenger RNAs is encoded into proteins. Here, we present an optimized protocol for assessing the translation rate in mouse adult microglia and cultured bone-marrow-derived macrophages. We describe steps for isolating cells, treating them with a puromycin-analog probe, and fluorescently labeling the puromycylated-polypeptide chains. We then detail their quantification by flow cytometry or with a fluorescent plate reader.For complete details on the use and execution of this protocol, please refer to Keane et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-12T00:42:20Z |
| format | Article |
| id | doaj.art-8c98f1f6414541d39a06571907cf0bfe |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-12T00:42:20Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-8c98f1f6414541d39a06571907cf0bfe2023-09-15T04:40:25ZengElsevierSTAR Protocols2666-16672023-12-0144102559Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophagesIgnazio Antignano0Lily Keane1Melania Capasso2Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), 53127 Bonn, Germany; Corresponding authorDeutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), 53127 Bonn, GermanyDeutsches Zentrum für Neurodegenerative Erkrankungen (DZNE), 53127 Bonn, Germany; Corresponding authorSummary: Protein synthesis, or mRNA translation, is the biological process through which genetic information stored in messenger RNAs is encoded into proteins. Here, we present an optimized protocol for assessing the translation rate in mouse adult microglia and cultured bone-marrow-derived macrophages. We describe steps for isolating cells, treating them with a puromycin-analog probe, and fluorescently labeling the puromycylated-polypeptide chains. We then detail their quantification by flow cytometry or with a fluorescent plate reader.For complete details on the use and execution of this protocol, please refer to Keane et al. (2021).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/S2666166723005269Flow Cytometry/Mass CytometryImmunologyNeuroscience |
| spellingShingle | Ignazio Antignano Lily Keane Melania Capasso Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages STAR Protocols Flow Cytometry/Mass Cytometry Immunology Neuroscience |
| title | Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages |
| title_full | Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages |
| title_fullStr | Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages |
| title_full_unstemmed | Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages |
| title_short | Assessing mRNA translation in mouse adult microglia and bone-marrow-derived macrophages |
| title_sort | assessing mrna translation in mouse adult microglia and bone marrow derived macrophages |
| topic | Flow Cytometry/Mass Cytometry Immunology Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723005269 |
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