Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns
This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the r...
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Veröffentlicht in: | Analytical biochemistry 1987-11, Vol.166 (2), p.267-275 |
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description | This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the risk of unstacking, columns packed with cellulose permit conventional elution of the protein zones and the use of a counter flow (to increase the effective length of the bed). For the same reason, electroosmotic flow is less disturbing. A continuous elution-migration technique adapted to suit the special requirements of displacement electrophoresis gave better separation than was obtainable by conventional elution. Normal human serum and a fresh hemolysate from human erythrocytes were used as samples. An expression for the volume velocity of the boundaries is derived. This parameter can be used to determine the maximum duration of a run and a suitable pump speed when continuous elution or a counter flow is employed. The special advantages of displacement electrophoresis in cellulose beds are discussed as well as general disadvantages of the displacement technique, including the risk that proteins precipitate during a run. |
doi_str_mv | 10.1016/0003-2697(87)90574-4 |
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Cellulose has virtually no molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the risk of unstacking, columns packed with cellulose permit conventional elution of the protein zones and the use of a counter flow (to increase the effective length of the bed). For the same reason, electroosmotic flow is less disturbing. A continuous elution-migration technique adapted to suit the special requirements of displacement electrophoresis gave better separation than was obtainable by conventional elution. Normal human serum and a fresh hemolysate from human erythrocytes were used as samples. An expression for the volume velocity of the boundaries is derived. This parameter can be used to determine the maximum duration of a run and a suitable pump speed when continuous elution or a counter flow is employed. The special advantages of displacement electrophoresis in cellulose beds are discussed as well as general disadvantages of the displacement technique, including the risk that proteins precipitate during a run.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/0003-2697(87)90574-4</identifier><identifier>PMID: 3434773</identifier><identifier>CODEN: ANBCA2</identifier><language>eng</language><publisher>San Diego, CA: Elsevier Inc</publisher><subject>Analytical biochemistry: general aspects, technics, instrumentation ; Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Blood Protein Electrophoresis ; Cellulose ; cellulose powder column ; continuous elution ; counter flow ; displacement electrophoresis ; Electrophoresis - instrumentation ; Electrophoresis, Polyacrylamide Gel ; Erythrocytes - analysis ; Fundamental and applied biological sciences. Psychology ; gel electrophoresis ; isotachophoresis ; protein aggregation ; proteins ; Proteins - analysis ; spacer techniques</subject><ispartof>Analytical biochemistry, 1987-11, Vol.166 (2), p.267-275</ispartof><rights>1987</rights><rights>1988 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c417t-e187246076336fdeb2f649da58965f09e7fa38b5e275122b5d54a01cae9943ff3</citedby><cites>FETCH-LOGICAL-c417t-e187246076336fdeb2f649da58965f09e7fa38b5e275122b5d54a01cae9943ff3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/0003269787905744$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7831079$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3434773$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Johansson, G.</creatorcontrib><creatorcontrib>Öfverstedt, L.-G.</creatorcontrib><creatorcontrib>Hjertén, S.</creatorcontrib><title>Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the risk of unstacking, columns packed with cellulose permit conventional elution of the protein zones and the use of a counter flow (to increase the effective length of the bed). For the same reason, electroosmotic flow is less disturbing. A continuous elution-migration technique adapted to suit the special requirements of displacement electrophoresis gave better separation than was obtainable by conventional elution. Normal human serum and a fresh hemolysate from human erythrocytes were used as samples. An expression for the volume velocity of the boundaries is derived. This parameter can be used to determine the maximum duration of a run and a suitable pump speed when continuous elution or a counter flow is employed. The special advantages of displacement electrophoresis in cellulose beds are discussed as well as general disadvantages of the displacement technique, including the risk that proteins precipitate during a run.</description><subject>Analytical biochemistry: general aspects, technics, instrumentation</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Blood Protein Electrophoresis</subject><subject>Cellulose</subject><subject>cellulose powder column</subject><subject>continuous elution</subject><subject>counter flow</subject><subject>displacement electrophoresis</subject><subject>Electrophoresis - instrumentation</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Erythrocytes - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>gel electrophoresis</subject><subject>isotachophoresis</subject><subject>protein aggregation</subject><subject>proteins</subject><subject>Proteins - analysis</subject><subject>spacer techniques</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkcFq3DAQhkVoSTabvkEKPpSQHJxqLFmyLoUSmqYQaA7NrSC08oio2JajsQN9-3i7yx6b08DMN8PPN4ydA78GDuoz51yUlTL6stFXhtdalvKIrYAbVXLBzTu2OiAn7JToD-cAslbH7FhIIbUWK_b7IePospviCxZtpLFzHnscpgI79FNO41PKSJGKy0hpcv7p0LkqUijGnCaMAxVpKDx23dwlwsKnbu4HOmPvg-sIP-zrmj3efvt1c1fe__z-4-brfekl6KlEaHQlFddKCBVa3FRBSdO6ujGqDtygDk40mxorXUNVbeq2lo6Dd2iMFCGINbvY3V3SPM9Ik-0jbdO4AdNMVmsjGgnNmyBIA0oCLKDcgT4noozBjjn2Lv-1wO3Wvt2qtVu1ttH2n30rl7WP-_vzpsf2sLTXvcw_7eeOvOtCdoOPdMB0I4AvWdfsyw7DRdpLxGzJRxw8tjEvT7Ftiv_P8QqNl6Gg</recordid><startdate>19871101</startdate><enddate>19871101</enddate><creator>Johansson, G.</creator><creator>Öfverstedt, L.-G.</creator><creator>Hjertén, S.</creator><general>Elsevier Inc</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>7QL</scope><scope>C1K</scope><scope>7X8</scope></search><sort><creationdate>19871101</creationdate><title>Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns</title><author>Johansson, G. ; Öfverstedt, L.-G. ; Hjertén, S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c417t-e187246076336fdeb2f649da58965f09e7fa38b5e275122b5d54a01cae9943ff3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>Analytical biochemistry: general aspects, technics, instrumentation</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Blood Protein Electrophoresis</topic><topic>Cellulose</topic><topic>cellulose powder column</topic><topic>continuous elution</topic><topic>counter flow</topic><topic>displacement electrophoresis</topic><topic>Electrophoresis - instrumentation</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Erythrocytes - analysis</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>gel electrophoresis</topic><topic>isotachophoresis</topic><topic>protein aggregation</topic><topic>proteins</topic><topic>Proteins - analysis</topic><topic>spacer techniques</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Johansson, G.</creatorcontrib><creatorcontrib>Öfverstedt, L.-G.</creatorcontrib><creatorcontrib>Hjertén, S.</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Johansson, G.</au><au>Öfverstedt, L.-G.</au><au>Hjertén, S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>1987-11-01</date><risdate>1987</risdate><volume>166</volume><issue>2</issue><spage>267</spage><epage>275</epage><pages>267-275</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><coden>ANBCA2</coden><abstract>This paper describes the separation of proteins by displacement electrophoresis on columns packed with cellulose powder as a stabilizing medium. Cellulose has virtually no molecularsieving properties and thus differs from dextran, polyacrylamide, and agarose in this respect. Therefore, without the risk of unstacking, columns packed with cellulose permit conventional elution of the protein zones and the use of a counter flow (to increase the effective length of the bed). For the same reason, electroosmotic flow is less disturbing. A continuous elution-migration technique adapted to suit the special requirements of displacement electrophoresis gave better separation than was obtainable by conventional elution. Normal human serum and a fresh hemolysate from human erythrocytes were used as samples. An expression for the volume velocity of the boundaries is derived. This parameter can be used to determine the maximum duration of a run and a suitable pump speed when continuous elution or a counter flow is employed. The special advantages of displacement electrophoresis in cellulose beds are discussed as well as general disadvantages of the displacement technique, including the risk that proteins precipitate during a run.</abstract><cop>San Diego, CA</cop><pub>Elsevier Inc</pub><pmid>3434773</pmid><doi>10.1016/0003-2697(87)90574-4</doi><tpages>9</tpages></addata></record> |
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subjects | Analytical biochemistry: general aspects, technics, instrumentation Analytical, structural and metabolic biochemistry Biological and medical sciences Blood Protein Electrophoresis Cellulose cellulose powder column continuous elution counter flow displacement electrophoresis Electrophoresis - instrumentation Electrophoresis, Polyacrylamide Gel Erythrocytes - analysis Fundamental and applied biological sciences. Psychology gel electrophoresis isotachophoresis protein aggregation proteins Proteins - analysis spacer techniques |
title | Preparative displacement electrophoresis (isotachophoresis) of proteins on cellulose columns |
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