Low-speed purification of human placental nuclei
A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteo...
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| Veröffentlicht in: | Placenta (Eastbourne) 1984-11, Vol.5 (6), p.523-532 |
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| container_title | Placenta (Eastbourne) |
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| creator | Resendez-Perez, D. Barrera-Saldaña, H.A. Morales-Vallarta, M.R. Ramirez-Bon, E. Leal-Garza, C.H. Feria-Velazco, A. Sanchez-Anzaldo, F.J. |
| description | A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei. |
| doi_str_mv | 10.1016/S0143-4004(84)80006-5 |
| format | Article |
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Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei.</description><identifier>ISSN: 0143-4004</identifier><identifier>EISSN: 1532-3102</identifier><identifier>DOI: 10.1016/S0143-4004(84)80006-5</identifier><identifier>PMID: 6527984</identifier><identifier>CODEN: PLACDF</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Alkaline Phosphatase - metabolism ; Biological and medical sciences ; Cell Fractionation - methods ; Cell Nucleus - enzymology ; Cell Nucleus - ultrastructure ; Centrifugation, Density Gradient ; DNA-Directed RNA Polymerases - metabolism ; Female ; Fundamental and applied biological sciences. Psychology ; Humans ; Mother. Fetoplacental unit. Mammary gland. Milk ; Placenta - ultrastructure ; Pregnancy ; Pregnancy. Parturition. Lactation ; Vertebrates: reproduction</subject><ispartof>Placenta (Eastbourne), 1984-11, Vol.5 (6), p.523-532</ispartof><rights>1985 W. B. Saunders Company Ltd.</rights><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c389t-204125d0d4e012a028142b439f5ff8cbc9dc2ea52b3091f52bbefe7b9dd17c533</citedby><cites>FETCH-LOGICAL-c389t-204125d0d4e012a028142b439f5ff8cbc9dc2ea52b3091f52bbefe7b9dd17c533</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0143400484800065$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65534</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9218850$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6527984$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Resendez-Perez, D.</creatorcontrib><creatorcontrib>Barrera-Saldaña, H.A.</creatorcontrib><creatorcontrib>Morales-Vallarta, M.R.</creatorcontrib><creatorcontrib>Ramirez-Bon, E.</creatorcontrib><creatorcontrib>Leal-Garza, C.H.</creatorcontrib><creatorcontrib>Feria-Velazco, A.</creatorcontrib><creatorcontrib>Sanchez-Anzaldo, F.J.</creatorcontrib><title>Low-speed purification of human placental nuclei</title><title>Placenta (Eastbourne)</title><addtitle>Placenta</addtitle><description>A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei.</description><subject>Alkaline Phosphatase - metabolism</subject><subject>Biological and medical sciences</subject><subject>Cell Fractionation - methods</subject><subject>Cell Nucleus - enzymology</subject><subject>Cell Nucleus - ultrastructure</subject><subject>Centrifugation, Density Gradient</subject><subject>DNA-Directed RNA Polymerases - metabolism</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Mother. Fetoplacental unit. Mammary gland. Milk</subject><subject>Placenta - ultrastructure</subject><subject>Pregnancy</subject><subject>Pregnancy. Parturition. Lactation</subject><subject>Vertebrates: reproduction</subject><issn>0143-4004</issn><issn>1532-3102</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1984</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1r3DAQhkVI2G62_QkBH0poDk5HsrSWTiUsSVtYyKHtWcjSiCj4q5KdkH9fbdbstTDwHuaZDx5CrijcUqDbr7-A8qrkAPyL5DcSALalOCNrKipWVhTYOVmfkA_kMqXnzChO2YqstoLVSvI1gf3wWqYR0RXjHIMP1kxh6IvBF09zZ_pibI3FfjJt0c-2xfCRXHjTJvy05Ib8ebj_vftR7h-__9zd7UtbSTWVDPIh4cBxBMoMMEk5a3ilvPBe2sYqZxkawZoKFPU5G_RYN8o5WltRVRtyfdw7xuHvjGnSXUgW29b0OMxJ10Kqmm5pBsURtHFIKaLXYwydiW-agj6Y0u-m9EGDlrkOprTIc1fLgbnp0J2mFjW5_3npm2RN66PpbUgnTDEqpYCMfTtimGW8BIw62YC9RRci2km7IfznkX9cKoPL</recordid><startdate>198411</startdate><enddate>198411</enddate><creator>Resendez-Perez, D.</creator><creator>Barrera-Saldaña, H.A.</creator><creator>Morales-Vallarta, M.R.</creator><creator>Ramirez-Bon, E.</creator><creator>Leal-Garza, C.H.</creator><creator>Feria-Velazco, A.</creator><creator>Sanchez-Anzaldo, F.J.</creator><general>Elsevier Ltd</general><general>Elsevier</general><scope>IQODW</scope><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></search><sort><creationdate>198411</creationdate><title>Low-speed purification of human placental nuclei</title><author>Resendez-Perez, D. ; Barrera-Saldaña, H.A. ; Morales-Vallarta, M.R. ; Ramirez-Bon, E. ; Leal-Garza, C.H. ; Feria-Velazco, A. ; Sanchez-Anzaldo, F.J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c389t-204125d0d4e012a028142b439f5ff8cbc9dc2ea52b3091f52bbefe7b9dd17c533</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1984</creationdate><topic>Alkaline Phosphatase - metabolism</topic><topic>Biological and medical sciences</topic><topic>Cell Fractionation - methods</topic><topic>Cell Nucleus - enzymology</topic><topic>Cell Nucleus - ultrastructure</topic><topic>Centrifugation, Density Gradient</topic><topic>DNA-Directed RNA Polymerases - metabolism</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Mother. Fetoplacental unit. Mammary gland. Milk</topic><topic>Placenta - ultrastructure</topic><topic>Pregnancy</topic><topic>Pregnancy. Parturition. Lactation</topic><topic>Vertebrates: reproduction</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Resendez-Perez, D.</creatorcontrib><creatorcontrib>Barrera-Saldaña, H.A.</creatorcontrib><creatorcontrib>Morales-Vallarta, M.R.</creatorcontrib><creatorcontrib>Ramirez-Bon, E.</creatorcontrib><creatorcontrib>Leal-Garza, C.H.</creatorcontrib><creatorcontrib>Feria-Velazco, A.</creatorcontrib><creatorcontrib>Sanchez-Anzaldo, F.J.</creatorcontrib><collection>Pascal-Francis</collection><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><jtitle>Placenta (Eastbourne)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Resendez-Perez, D.</au><au>Barrera-Saldaña, H.A.</au><au>Morales-Vallarta, M.R.</au><au>Ramirez-Bon, E.</au><au>Leal-Garza, C.H.</au><au>Feria-Velazco, A.</au><au>Sanchez-Anzaldo, F.J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Low-speed purification of human placental nuclei</atitle><jtitle>Placenta (Eastbourne)</jtitle><addtitle>Placenta</addtitle><date>1984-11</date><risdate>1984</risdate><volume>5</volume><issue>6</issue><spage>523</spage><epage>532</epage><pages>523-532</pages><issn>0143-4004</issn><eissn>1532-3102</eissn><coden>PLACDF</coden><abstract>A simple method for the purification of human placental nuclei is described. Nuclei were isolated by homogenizing tissue in standard saline citrate solution in the presence of zinc chloride to stabilize the nuclear membranes, NP40 as non-ionic detergent and sodium bisulphite for inhibition of proteolytic activity. Nuclei purification was achieved by low-speed centrifugation through a discontinuous sucrose gradient. The purified nuclei were evaluated by morphological criteria using phase contrast and electron microscopy. The extent of contamination by cytoplasmic debris was estimated by Papanicolaou's staining technique. Biochemical criteria include measurements of alkaline phosphatase activity as a plasma membrane enzyme marker and DNA-dependent RNA polymerase activity for the functional integrity of nuclear components. Transcriptionally active nuclei were obtained but the yield of nuclei was low; however, this low yield is compensated by the high degree of purity, the simplicity of the method and the functional and morphological integrity of the purified nuclei.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>6527984</pmid><doi>10.1016/S0143-4004(84)80006-5</doi><tpages>10</tpages></addata></record> |
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| ispartof | Placenta (Eastbourne), 1984-11, Vol.5 (6), p.523-532 |
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| source | MEDLINE; Elsevier ScienceDirect Journals Complete |
| subjects | Alkaline Phosphatase - metabolism Biological and medical sciences Cell Fractionation - methods Cell Nucleus - enzymology Cell Nucleus - ultrastructure Centrifugation, Density Gradient DNA-Directed RNA Polymerases - metabolism Female Fundamental and applied biological sciences. Psychology Humans Mother. Fetoplacental unit. Mammary gland. Milk Placenta - ultrastructure Pregnancy Pregnancy. Parturition. Lactation Vertebrates: reproduction |
| title | Low-speed purification of human placental nuclei |
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