2-D differential membrane proteome analysis of scarce protein samples
Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) w...
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| Veröffentlicht in: | Proteomics (Weinheim) 2006-08, Vol.6 (16), p.4506-4513 |
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| creator | Helling, Stefan Schmitt, Edgar Joppich, Cornelia Schulenborg, Thomas Müllner, Stefan Felske-Müller, Stephanie Wiebringhaus, Thomas Becker, Gabriele Linsenmann, Gudrun Sitek, Barbara Lutter, Petra Meyer, Helmut E. Marcus, Katrin |
| description | Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X‐114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D‐CTAB/SDS‐PAGE. For a differential analysis 3 μg protein was found to be sufficient to detect proteins in a widespread well‐separated diagonal spot pattern. |
| doi_str_mv | 10.1002/pmic.200600169 |
| format | Article |
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In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X‐114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D‐CTAB/SDS‐PAGE. 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Psychology ; Humans ; Membrane proteins ; Membrane Proteins - analysis ; Membrane Proteins - isolation & purification ; Mice ; Miscellaneous ; Molecular Sequence Data ; Multiplexed proteomics technology ; Proteins ; Proteome ; Proteomics methods ; satDIGE ; Spectrometry, Mass, Electrospray Ionization</subject><ispartof>Proteomics (Weinheim), 2006-08, Vol.6 (16), p.4506-4513</ispartof><rights>Copyright © 2006 WILEY‐VCH Verlag GmbH & Co. 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In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X‐114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D‐CTAB/SDS‐PAGE. For a differential analysis 3 μg protein was found to be sufficient to detect proteins in a widespread well‐separated diagonal spot pattern.</description><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Fluorescence two-dimensional difference gel electrophoresis</subject><subject>Fluorescent Dyes - chemistry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Membrane proteins</subject><subject>Membrane Proteins - analysis</subject><subject>Membrane Proteins - isolation & purification</subject><subject>Mice</subject><subject>Miscellaneous</subject><subject>Molecular Sequence Data</subject><subject>Multiplexed proteomics technology</subject><subject>Proteins</subject><subject>Proteome</subject><subject>Proteomics methods</subject><subject>satDIGE</subject><subject>Spectrometry, Mass, Electrospray Ionization</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtPGzEURi1UBJSyZYlm0-4m-DF-3GUVIEWiPKRWZWd5PNeS25lMaidq8-9rlFHojpUt-3z3uzqEnDM6Y5Tyy9UQ_YxTqihlCg7ICVNM1mAUe7e_S3FM3uf8syDagD4ix0wZIcv7Cbnm9VXVxRAw4XIdXV8NOLTJLbFapXGN44CVW7p-m2OuxlBl75Kf_uKyym5Y9Zg_kMPg-oxn03lKvt9cf5t_qe8eFrfzz3e1bxoONSIGLkG30mgtAZmB1gFFLmknOBPBe80AONCgHDKpOxAd94475WQLIE7Jp93c0v97g3lth5g99n3Zd9xkq4w2jTTiTZBBo0oVK-BsB_o05pww2FWKg0tby6h9MWxfDNu94RK4mCZv2gG7V3xSWoCPE-CKrD4Ulz7mV85QQRveFA523J_Y4_aNWvv49Xb-_xL1LhvzGv_usy79skoLLe2P-4V9BsbV4obaJ_EPbr-i-w</recordid><startdate>20060801</startdate><enddate>20060801</enddate><creator>Helling, Stefan</creator><creator>Schmitt, Edgar</creator><creator>Joppich, Cornelia</creator><creator>Schulenborg, Thomas</creator><creator>Müllner, Stefan</creator><creator>Felske-Müller, Stephanie</creator><creator>Wiebringhaus, Thomas</creator><creator>Becker, Gabriele</creator><creator>Linsenmann, Gudrun</creator><creator>Sitek, Barbara</creator><creator>Lutter, Petra</creator><creator>Meyer, Helmut E.</creator><creator>Marcus, Katrin</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><general>Wiley-VCH</general><scope>BSCLL</scope><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>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20060801</creationdate><title>2-D differential membrane proteome analysis of scarce protein samples</title><author>Helling, Stefan ; Schmitt, Edgar ; Joppich, Cornelia ; Schulenborg, Thomas ; Müllner, Stefan ; Felske-Müller, Stephanie ; Wiebringhaus, Thomas ; Becker, Gabriele ; Linsenmann, Gudrun ; Sitek, Barbara ; Lutter, Petra ; Meyer, Helmut E. ; Marcus, Katrin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4429-eeef2597b587759e189ba90e250d3213fcc7199290f6ae157d93d2ca2a6a5b993</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Fluorescence two-dimensional difference gel electrophoresis</topic><topic>Fluorescent Dyes - chemistry</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Membrane proteins</topic><topic>Membrane Proteins - analysis</topic><topic>Membrane Proteins - isolation & purification</topic><topic>Mice</topic><topic>Miscellaneous</topic><topic>Molecular Sequence Data</topic><topic>Multiplexed proteomics technology</topic><topic>Proteins</topic><topic>Proteome</topic><topic>Proteomics methods</topic><topic>satDIGE</topic><topic>Spectrometry, Mass, Electrospray Ionization</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Helling, Stefan</creatorcontrib><creatorcontrib>Schmitt, Edgar</creatorcontrib><creatorcontrib>Joppich, Cornelia</creatorcontrib><creatorcontrib>Schulenborg, Thomas</creatorcontrib><creatorcontrib>Müllner, Stefan</creatorcontrib><creatorcontrib>Felske-Müller, Stephanie</creatorcontrib><creatorcontrib>Wiebringhaus, Thomas</creatorcontrib><creatorcontrib>Becker, Gabriele</creatorcontrib><creatorcontrib>Linsenmann, Gudrun</creatorcontrib><creatorcontrib>Sitek, Barbara</creatorcontrib><creatorcontrib>Lutter, Petra</creatorcontrib><creatorcontrib>Meyer, Helmut E.</creatorcontrib><creatorcontrib>Marcus, Katrin</creatorcontrib><collection>Istex</collection><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>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Helling, Stefan</au><au>Schmitt, Edgar</au><au>Joppich, Cornelia</au><au>Schulenborg, Thomas</au><au>Müllner, Stefan</au><au>Felske-Müller, Stephanie</au><au>Wiebringhaus, Thomas</au><au>Becker, Gabriele</au><au>Linsenmann, Gudrun</au><au>Sitek, Barbara</au><au>Lutter, Petra</au><au>Meyer, Helmut E.</au><au>Marcus, Katrin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>2-D differential membrane proteome analysis of scarce protein samples</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2006-08-01</date><risdate>2006</risdate><volume>6</volume><issue>16</issue><spage>4506</spage><epage>4513</epage><pages>4506-4513</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>Proteome studies with small sample amounts are difficult to perform, especially when membrane proteins are the focus of interest. In our study a new method for the analysis of scarce membrane protein samples combining large gel 2‐D‐CTAB/SDS‐PAGE with fluorescence dye saturation labelling (satDIGE) was developed, allowing a highly sensitive differential analysis of different cell states. After Triton X‐114 phase partitioning, enriched membrane protein samples of T cells were labelled at cysteine residues using fluorescence dyes and separated by large gel 2D‐CTAB/SDS‐PAGE. For a differential analysis 3 μg protein was found to be sufficient to detect proteins in a widespread well‐separated diagonal spot pattern.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>16835853</pmid><doi>10.1002/pmic.200600169</doi><tpages>8</tpages></addata></record> |
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| subjects | Amino Acid Sequence Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Cells, Cultured Electrophoresis, Gel, Two-Dimensional Fluorescence two-dimensional difference gel electrophoresis Fluorescent Dyes - chemistry Fundamental and applied biological sciences. Psychology Humans Membrane proteins Membrane Proteins - analysis Membrane Proteins - isolation & purification Mice Miscellaneous Molecular Sequence Data Multiplexed proteomics technology Proteins Proteome Proteomics methods satDIGE Spectrometry, Mass, Electrospray Ionization |
| title | 2-D differential membrane proteome analysis of scarce protein samples |
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