Procedure for the isolation of mitochondria, cytosolic and nuclear material from a single piece of neurological tissue for high-throughput mass spectral analysis
The isolation of high-purity cellular biomacromolecules and sub-cellular organelles is an essential aspect to mass spectrometry based studies. Mitochondria are sub-cellular organelles that perform a central role in cellular energy production. Mitochondria are of great interest due to their potential...
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| Veröffentlicht in: | Journal of neuroscience methods 2011-04, Vol.197 (2), p.279-282 |
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| container_end_page | 282 |
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| container_title | Journal of neuroscience methods |
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| creator | Timmons, Michael D. Bradley, Melissa A. Lovell, Mark A. Lynn, Bert C. |
| description | The isolation of high-purity cellular biomacromolecules and sub-cellular organelles is an essential aspect to mass spectrometry based studies. Mitochondria are sub-cellular organelles that perform a central role in cellular energy production. Mitochondria are of great interest due to their potential to generate reactive oxygen species (ROS) and susceptibility to oxidative damage and subsequent functional impairment. Current methods of mitochondria isolation are optimized for respiratory-based studies that favor viability. Whereas, proteomic and lipidomics studies of mitochondria require procedures that optimize for purity and enrichment. We describe a procedure derived from previously established methods for the isolation of mitochondria, nuclear and cytosolic fractions from a neurological tissue sample. In addition to the isolation being of significant purity for mass spectral based ‘-omics’ analysis, mitochondrial yields were routinely 500μg per tissue wet weight, allowing multiple studies to be conducted from a single isolation procedure. |
| doi_str_mv | 10.1016/j.jneumeth.2011.02.027 |
| format | Article |
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All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c399t-434494d8c54d2343395d1f6fb74ce5b4ea15737d8aa2c8a50523cab5bd785da43</citedby><cites>FETCH-LOGICAL-c399t-434494d8c54d2343395d1f6fb74ce5b4ea15737d8aa2c8a50523cab5bd785da43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0165027011001221$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21392528$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Timmons, Michael D.</creatorcontrib><creatorcontrib>Bradley, Melissa A.</creatorcontrib><creatorcontrib>Lovell, Mark A.</creatorcontrib><creatorcontrib>Lynn, Bert C.</creatorcontrib><title>Procedure for the isolation of mitochondria, cytosolic and nuclear material from a single piece of neurological tissue for high-throughput mass spectral analysis</title><title>Journal of neuroscience methods</title><addtitle>J Neurosci Methods</addtitle><description>The isolation of high-purity cellular biomacromolecules and sub-cellular organelles is an essential aspect to mass spectrometry based studies. Mitochondria are sub-cellular organelles that perform a central role in cellular energy production. Mitochondria are of great interest due to their potential to generate reactive oxygen species (ROS) and susceptibility to oxidative damage and subsequent functional impairment. Current methods of mitochondria isolation are optimized for respiratory-based studies that favor viability. Whereas, proteomic and lipidomics studies of mitochondria require procedures that optimize for purity and enrichment. We describe a procedure derived from previously established methods for the isolation of mitochondria, nuclear and cytosolic fractions from a neurological tissue sample. In addition to the isolation being of significant purity for mass spectral based ‘-omics’ analysis, mitochondrial yields were routinely 500μg per tissue wet weight, allowing multiple studies to be conducted from a single isolation procedure.</description><subject>Cell Fractionation - methods</subject><subject>Cell Nucleus - chemistry</subject><subject>Cell Nucleus - ultrastructure</subject><subject>Cytosol - chemistry</subject><subject>Cytosol - ultrastructure</subject><subject>Genomics</subject><subject>Humans</subject><subject>Isolation procedure</subject><subject>Lipidomics</subject><subject>Mass Spectrometry - methods</subject><subject>Mitochondria</subject><subject>Mitochondria - chemistry</subject><subject>Mitochondria - ultrastructure</subject><subject>Nervous System Diseases - metabolism</subject><subject>Nervous System Diseases - pathology</subject><subject>Neurological tissue</subject><subject>Percoll</subject><subject>Subcellular Fractions - chemistry</subject><subject>Subcellular Fractions - ultrastructure</subject><issn>0165-0270</issn><issn>1872-678X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkd2KFDEQhYMo7rj6CkvuvLHH_HZ37pRl_YEFvVDwLqST6ukM6U6bpIV5HN_UDLPrrUJBoOqrc0IdhG4o2VNC27fH_XGBbYYy7RmhdE9Yre4J2tG-Y03b9T-eol0FZVPb5Aq9yPlICBGKtM_RFaNcMcn6Hfr9NUULbkuAx5hwmQD7HIMpPi44jnj2JdopLi558wbbU4l16i02i8PLZgOYhGdToI4DHlOcscHZL4cAePVg4axRP5piiAdvK1N8ztvFbPKHqSlTitthWrdSdXLGeQVbUgXNYsIp-_wSPRtNyPDq4b1G3z_cfbv91Nx_-fj59v19Y7lSpRFcCCVcb6VwjAvOlXR0bMehExbkIMBQ2fHO9cYw2xtJJOPWDHJwXS-dEfwavb7orin-3CAXPftsIQSzQNyy7luhWqLa7j_I6k87RSvZXkibYs4JRr0mP5t00pToc476qB9z1OccNWG1zhY3DxbbMIP7u_YYXAXeXQCoJ_nlIelsPSw1Sp_qAbWL_l8efwCRcLaU</recordid><startdate>20110430</startdate><enddate>20110430</enddate><creator>Timmons, Michael D.</creator><creator>Bradley, Melissa A.</creator><creator>Lovell, Mark A.</creator><creator>Lynn, Bert C.</creator><general>Elsevier B.V</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>7TK</scope></search><sort><creationdate>20110430</creationdate><title>Procedure for the isolation of mitochondria, cytosolic and nuclear material from a single piece of neurological tissue for high-throughput mass spectral analysis</title><author>Timmons, Michael D. ; 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| fulltext | fulltext |
| identifier | ISSN: 0165-0270 |
| ispartof | Journal of neuroscience methods, 2011-04, Vol.197 (2), p.279-282 |
| issn | 0165-0270 1872-678X |
| language | eng |
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| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Cell Fractionation - methods Cell Nucleus - chemistry Cell Nucleus - ultrastructure Cytosol - chemistry Cytosol - ultrastructure Genomics Humans Isolation procedure Lipidomics Mass Spectrometry - methods Mitochondria Mitochondria - chemistry Mitochondria - ultrastructure Nervous System Diseases - metabolism Nervous System Diseases - pathology Neurological tissue Percoll Subcellular Fractions - chemistry Subcellular Fractions - ultrastructure |
| title | Procedure for the isolation of mitochondria, cytosolic and nuclear material from a single piece of neurological tissue for high-throughput mass spectral analysis |
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