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Article: Subcellular Fractionation of Cultured Human Cell Lines

TitleSubcellular Fractionation of Cultured Human Cell Lines
Authors
Issue Date2013
PublisherBio-protocol. The Journal's web site is located at http://www.bio-protocol.org/Default.aspx
Citation
Bio-protocol, 2013, v. 3 n. 9, article no. e754 How to Cite?
AbstractSubcellular localization is crucial for the proper functioning of a protein. Deregulation of subcellular localization may lead to pathological consequences and result in diseases like cancer. Immuno-fluorescent staining and subcellular fractionation can be used to determine localization of a protein. Here we discuss a protocol to separate the nuclear, cytosolic, and membrane fractions of cultured human cell lines using a centrifuge and ultracentrifuge. The membrane fraction contains plasma membranes and ER-golgi membranes, but no mitochondria or nuclear structures. The fractions can be further analyzed using Western blotting. This protocol is based on that from Dr. Richard Patten at Abcam (1), and was modified and utilized in a publication by Huang et al. (2012).
Persistent Identifierhttp://hdl.handle.net/10722/202755
ISSN

 

DC FieldValueLanguage
dc.contributor.authorYu, Zen_US
dc.contributor.authorHuang, Zen_US
dc.contributor.authorLung, ML-
dc.date.accessioned2014-09-19T09:33:36Z-
dc.date.available2014-09-19T09:33:36Z-
dc.date.issued2013en_US
dc.identifier.citationBio-protocol, 2013, v. 3 n. 9, article no. e754en_US
dc.identifier.issn2331-8325-
dc.identifier.urihttp://hdl.handle.net/10722/202755-
dc.description.abstractSubcellular localization is crucial for the proper functioning of a protein. Deregulation of subcellular localization may lead to pathological consequences and result in diseases like cancer. Immuno-fluorescent staining and subcellular fractionation can be used to determine localization of a protein. Here we discuss a protocol to separate the nuclear, cytosolic, and membrane fractions of cultured human cell lines using a centrifuge and ultracentrifuge. The membrane fraction contains plasma membranes and ER-golgi membranes, but no mitochondria or nuclear structures. The fractions can be further analyzed using Western blotting. This protocol is based on that from Dr. Richard Patten at Abcam (1), and was modified and utilized in a publication by Huang et al. (2012).-
dc.languageengen_US
dc.publisherBio-protocol. The Journal's web site is located at http://www.bio-protocol.org/Default.aspx-
dc.relation.ispartofBio-protocolen_US
dc.titleSubcellular Fractionation of Cultured Human Cell Linesen_US
dc.typeArticleen_US
dc.identifier.emailYu, Z: zvyu@hku.hken_US
dc.identifier.emailLung, ML: mlilung@hku.hken_US
dc.identifier.authorityLung, ML=rp00300en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.hkuros240230en_US
dc.identifier.volume3en_US
dc.identifier.issue9en_US
dc.publisher.placeUnited States-

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