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Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum
A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content protein...
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| Published in: | The Journal of cell biology 1982-04, Vol.93 (1), p.97-102 |
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| Main Authors: | , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c463t-b4b0f1225c3c54329c0148a9613b1bfd6228f933cd7ca1311d4a1c76122003983 |
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| container_end_page | 102 |
| container_issue | 1 |
| container_start_page | 97 |
| container_title | The Journal of cell biology |
| container_volume | 93 |
| creator | Fujiki, Yukio Hubbard, Ann L. Fowler, Stanley Lazarow, Paul B. |
| description | A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper the procedure is applied to peroxisomes and mitochondria. |
| doi_str_mv | 10.1083/jcb.93.1.97 |
| format | article |
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The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper the procedure is applied to peroxisomes and mitochondria.</description><identifier>ISSN: 0021-9525</identifier><identifier>EISSN: 1540-8140</identifier><identifier>DOI: 10.1083/jcb.93.1.97</identifier><identifier>PMID: 7068762</identifier><language>eng</language><publisher>United States: Rockefeller University Press</publisher><subject>Animals ; Carbonates ; Cell Fractionation - methods ; Cell membranes ; Centrifugation, Density Gradient - methods ; Cytochromes ; Female ; Intracellular Membranes - ultrastructure ; Liver ; Liver - ultrastructure ; Membrane proteins ; Microscopy, Electron ; Microsomes ; Microsomes, Liver - ultrastructure ; P branes ; Rats ; Rats, Inbred Strains ; Ribosomes ; RNA ; Sarcoplasmic Reticulum - ultrastructure ; Sodium</subject><ispartof>The Journal of cell biology, 1982-04, Vol.93 (1), p.97-102</ispartof><rights>Copyright 1982 The Rockefeller University Press</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c463t-b4b0f1225c3c54329c0148a9613b1bfd6228f933cd7ca1311d4a1c76122003983</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7068762$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Fujiki, Yukio</creatorcontrib><creatorcontrib>Hubbard, Ann L.</creatorcontrib><creatorcontrib>Fowler, Stanley</creatorcontrib><creatorcontrib>Lazarow, Paul B.</creatorcontrib><title>Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum</title><title>The Journal of cell biology</title><addtitle>J Cell Biol</addtitle><description>A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper the procedure is applied to peroxisomes and mitochondria.</description><subject>Animals</subject><subject>Carbonates</subject><subject>Cell Fractionation - methods</subject><subject>Cell membranes</subject><subject>Centrifugation, Density Gradient - methods</subject><subject>Cytochromes</subject><subject>Female</subject><subject>Intracellular Membranes - ultrastructure</subject><subject>Liver</subject><subject>Liver - ultrastructure</subject><subject>Membrane proteins</subject><subject>Microscopy, Electron</subject><subject>Microsomes</subject><subject>Microsomes, Liver - ultrastructure</subject><subject>P branes</subject><subject>Rats</subject><subject>Rats, Inbred Strains</subject><subject>Ribosomes</subject><subject>RNA</subject><subject>Sarcoplasmic Reticulum - ultrastructure</subject><subject>Sodium</subject><issn>0021-9525</issn><issn>1540-8140</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1982</creationdate><recordtype>article</recordtype><recordid>eNpVkdGLFSEYxSWK7bb11GuBT73E3PzUGcceguWy1YWNoLZnUccpL844q06w_31e7mUrEETOz6PfOQi9BLIF0rN3B2u2km1hK8UjtIGWk6YHTh6jDSEUGtnS9il6lvOBEMIFZxfoQpCuFx3doLt9jkEXH2ccR7yfS9LWhbAGnfAXN5mkZ5exua8HPecj8z0Ofp3wTicTZ10cvk1Ol8nN5T2-Wpbg7cmuRHw9D3EJOk_e4m-ueLuGdXqOnow6ZPfivF-iHx-vb3efm5uvn_a7q5vG8o6VxnBDRqC0tcy2nFFpCfBeyw6YATMOHaX9KBmzg7AaGMDANVjR1SuEMNmzS_Th5LusZnKDdcfZglqSn3S6V1F79b8y-1_qZ_ytKEBdrBq8ORukeLe6XNTk8zGcGklcsxK8ptt3bQXfnkCbYs7JjQ-PAFHHhlRtSEmmQElR6df__uuBPVdS9Vcn_ZBLTH-tOlKHEuwPoBSXeQ</recordid><startdate>19820401</startdate><enddate>19820401</enddate><creator>Fujiki, Yukio</creator><creator>Hubbard, Ann L.</creator><creator>Fowler, Stanley</creator><creator>Lazarow, Paul B.</creator><general>Rockefeller University Press</general><general>The Rockefeller University Press</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>5PM</scope></search><sort><creationdate>19820401</creationdate><title>Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum</title><author>Fujiki, Yukio ; Hubbard, Ann L. ; Fowler, Stanley ; Lazarow, Paul B.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c463t-b4b0f1225c3c54329c0148a9613b1bfd6228f933cd7ca1311d4a1c76122003983</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1982</creationdate><topic>Animals</topic><topic>Carbonates</topic><topic>Cell Fractionation - methods</topic><topic>Cell membranes</topic><topic>Centrifugation, Density Gradient - methods</topic><topic>Cytochromes</topic><topic>Female</topic><topic>Intracellular Membranes - ultrastructure</topic><topic>Liver</topic><topic>Liver - ultrastructure</topic><topic>Membrane proteins</topic><topic>Microscopy, Electron</topic><topic>Microsomes</topic><topic>Microsomes, Liver - ultrastructure</topic><topic>P branes</topic><topic>Rats</topic><topic>Rats, Inbred Strains</topic><topic>Ribosomes</topic><topic>RNA</topic><topic>Sarcoplasmic Reticulum - ultrastructure</topic><topic>Sodium</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Fujiki, Yukio</creatorcontrib><creatorcontrib>Hubbard, Ann L.</creatorcontrib><creatorcontrib>Fowler, Stanley</creatorcontrib><creatorcontrib>Lazarow, Paul B.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>The Journal of cell biology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Fujiki, Yukio</au><au>Hubbard, Ann L.</au><au>Fowler, Stanley</au><au>Lazarow, Paul B.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum</atitle><jtitle>The Journal of cell biology</jtitle><addtitle>J Cell Biol</addtitle><date>1982-04-01</date><risdate>1982</risdate><volume>93</volume><issue>1</issue><spage>97</spage><epage>102</epage><pages>97-102</pages><issn>0021-9525</issn><eissn>1540-8140</eissn><abstract>A rapid and simple method for the isolation of membranes from subcellular organelles is described. The procedure consists of diluting the organelles in ice-cold 100 mM Na2CO3followed by centrifugation to pellet the membranes. Closed vesicles are converted to open membrane sheets, and content proteins and peripheral membrane proteins are released in soluble form. Here we document the method by applying it to various subfractions of a rat liver microsomal fraction, prepared by continuous density gradient centrifugation according to Beaufay et al. The results confirm and extend those of previous investigators on the distribution of enzymes and proteins among the membranes of the smooth and rough endoplasmic reticulum. In the accompanying paper the procedure is applied to peroxisomes and mitochondria.</abstract><cop>United States</cop><pub>Rockefeller University Press</pub><pmid>7068762</pmid><doi>10.1083/jcb.93.1.97</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0021-9525 |
| ispartof | The Journal of cell biology, 1982-04, Vol.93 (1), p.97-102 |
| issn | 0021-9525 1540-8140 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_2112113 |
| source | Alma/SFX Local Collection |
| subjects | Animals Carbonates Cell Fractionation - methods Cell membranes Centrifugation, Density Gradient - methods Cytochromes Female Intracellular Membranes - ultrastructure Liver Liver - ultrastructure Membrane proteins Microscopy, Electron Microsomes Microsomes, Liver - ultrastructure P branes Rats Rats, Inbred Strains Ribosomes RNA Sarcoplasmic Reticulum - ultrastructure Sodium |
| title | Isolation of Intracellular Membranes by Means of Sodium Carbonate Treatment: Application to Endoplasmic Reticulum |
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