Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer
Summary: Efficient metabolism, or the means by which cells produce energy resources, is critical for proper effector function. Here, we present a protocol for examining the bioenergetics and mitochondrial fuel utilization of primary murine autoreactive immunocytes using cellular metabolism-modulatin...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2024-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166724001369 |
| _version_ | 1827307861920710656 |
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| author | Colleen L. Mayberry John J. Wilson Britney Sison Chih-Hao Chang |
| author_facet | Colleen L. Mayberry John J. Wilson Britney Sison Chih-Hao Chang |
| author_sort | Colleen L. Mayberry |
| collection | DOAJ |
| description | Summary: Efficient metabolism, or the means by which cells produce energy resources, is critical for proper effector function. Here, we present a protocol for examining the bioenergetics and mitochondrial fuel utilization of primary murine autoreactive immunocytes using cellular metabolism-modulating drugs. We describe steps for plate calibration, isolation of primary immunocytes, and Seahorse assay plate preparation. We then detail procedures for performing the XF Cell Mito Stress Test followed by bioenergetics calculations and statistics.For complete details on the use and execution of this protocol, please refer to Wilson et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-24T18:47:44Z |
| format | Article |
| id | doaj.art-b43702ddeede41e59e361ef86770ffa0 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-24T18:47:44Z |
| publishDate | 2024-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-b43702ddeede41e59e361ef86770ffa02024-03-27T04:52:51ZengElsevierSTAR Protocols2666-16672024-06-0152102971Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux AnalyzerColleen L. Mayberry0John J. Wilson1Britney Sison2Chih-Hao Chang3The Jackson Laboratory, Bar Harbor, ME 04609, USAThe Jackson Laboratory, Bar Harbor, ME 04609, USAThe Jackson Laboratory, Bar Harbor, ME 04609, USAThe Jackson Laboratory, Bar Harbor, ME 04609, USA; Graduate School of Biomedical Sciences and Engineering, The University of Maine, Orono, ME 04469, USA; Graduate School of Biomedical Sciences, Tufts University School of Medicine, Boston, MA 02111, USA; Corresponding authorSummary: Efficient metabolism, or the means by which cells produce energy resources, is critical for proper effector function. Here, we present a protocol for examining the bioenergetics and mitochondrial fuel utilization of primary murine autoreactive immunocytes using cellular metabolism-modulating drugs. We describe steps for plate calibration, isolation of primary immunocytes, and Seahorse assay plate preparation. We then detail procedures for performing the XF Cell Mito Stress Test followed by bioenergetics calculations and statistics.For complete details on the use and execution of this protocol, please refer to Wilson 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/S2666166724001369Cell-based AssaysImmunologyMetabolism |
| spellingShingle | Colleen L. Mayberry John J. Wilson Britney Sison Chih-Hao Chang Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer STAR Protocols Cell-based Assays Immunology Metabolism |
| title | Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer |
| title_full | Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer |
| title_fullStr | Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer |
| title_full_unstemmed | Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer |
| title_short | Protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the Seahorse Extracellular Flux Analyzer |
| title_sort | protocol to assess bioenergetics and mitochondrial fuel usage in murine autoreactive immunocytes using the seahorse extracellular flux analyzer |
| topic | Cell-based Assays Immunology Metabolism |
| url | http://www.sciencedirect.com/science/article/pii/S2666166724001369 |
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