Polyacrylamide lamination enables mass spectrometry compatible staining and in-gel digestion of proteins separated by agarose IEF
Agarose IEF enables the separation of large proteins and protein complexes. A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to sep...
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| Veröffentlicht in: | Proteomics (Weinheim) 2007-10, Vol.7 (19), p.3441-3444 |
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| creator | Hellman, Jukka |
| description | Agarose IEF enables the separation of large proteins and protein complexes. A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to separate the proteins, followed by layering the agarose with polyacrylamide. The formed laminate gels were seamless and durable and they were readily detached from the polyester. The gels were amenable to MS compatible staining. The sensitivity obtained with the acidic silver staining method was 20-50 ng/band of myoglobin. Laminated agarose was a suitable matrix for in-gel digestion based generation of tryptic peptides for MALDI-MS. |
| doi_str_mv | 10.1002/pmic.200700299 |
| format | Article |
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A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to separate the proteins, followed by layering the agarose with polyacrylamide. The formed laminate gels were seamless and durable and they were readily detached from the polyester. The gels were amenable to MS compatible staining. The sensitivity obtained with the acidic silver staining method was 20-50 ng/band of myoglobin. Laminated agarose was a suitable matrix for in-gel digestion based generation of tryptic peptides for MALDI-MS.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200700299</identifier><identifier>PMID: 17722206</identifier><language>eng</language><publisher>Weinheim: Wiley-VCH Verlag</publisher><subject>Acrylic Resins - chemistry ; Agarose IEF ; Amino Acid Sequence ; Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Electrophoresis, Agar Gel - methods ; Fundamental and applied biological sciences. 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A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to separate the proteins, followed by layering the agarose with polyacrylamide. The formed laminate gels were seamless and durable and they were readily detached from the polyester. The gels were amenable to MS compatible staining. The sensitivity obtained with the acidic silver staining method was 20-50 ng/band of myoglobin. Laminated agarose was a suitable matrix for in-gel digestion based generation of tryptic peptides for MALDI-MS.</description><subject>Acrylic Resins - chemistry</subject><subject>Agarose IEF</subject><subject>Amino Acid Sequence</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Electrophoresis, Agar Gel - methods</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GelBond</subject><subject>In-gel digestion</subject><subject>Isoelectric Focusing - methods</subject><subject>MALDI-MS</subject><subject>Mass Spectrometry - methods</subject><subject>Miscellaneous</subject><subject>Molecular Sequence Data</subject><subject>Proteins</subject><subject>Proteins - analysis</subject><subject>Proteins - genetics</subject><subject>Silver staining</subject><subject>Staining and Labeling - methods</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1v1DAQxSMEoqVw5Qi-wC2LHSd2ckSrfqzUQkUpcLMmzjgyJE5qZ0Vz7H-Ol6y23DiNLf_em-eXJK8ZXTFKsw9jb_Uqo1TGS1U9SY6ZYEValYI9PZwLfpS8COEnpUyWlXyeHDEpsyyj4jh5uB66GbSfO-htg2Q3HEx2cAQd1B0G0kMIJIyoJz_0OPmZ6KEfIxNfSZjAOutaAq4h1qUtdqSxLYa_FoMhox8mtC464AgeJmxIPRNowQ8Byeb07GXyzEAX8NV-niS3Z6df1xfp5efzzfrjZapzXlRp2ZiywCJjgjMmatrUFeRoqESqDdVM1zVnZSOBSRQmftMwhiC5EabJKyH4SfJ-8Y2J7rYxoOpt0Nh14HDYBiVKXsRaeARXC6hjxODRqNHbHvysGFW70tWudHUoPQre7J23dY_NI75vOQLv9gAEDZ3x4LQNj1wVKcZZ5KqF-207nP-zVl1fbdb_hkgXrQ0T3h-04H8pIbks1PdP52r9Rfy4-nZzocrIv114A4OC1sc8tze7GJSWrMxzwf8AKg22qw</recordid><startdate>20071001</startdate><enddate>20071001</enddate><creator>Hellman, Jukka</creator><general>Wiley-VCH Verlag</general><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><general>Wiley-VCH</general><scope>FBQ</scope><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>7X8</scope></search><sort><creationdate>20071001</creationdate><title>Polyacrylamide lamination enables mass spectrometry compatible staining and in-gel digestion of proteins separated by agarose IEF</title><author>Hellman, Jukka</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4359-8df85e52163116b0db9a4ef07e0cf0c1cbb318d7a17e6f897f11ea73f6fd49663</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Acrylic Resins - chemistry</topic><topic>Agarose IEF</topic><topic>Amino Acid Sequence</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Electrophoresis, Agar Gel - methods</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GelBond</topic><topic>In-gel digestion</topic><topic>Isoelectric Focusing - methods</topic><topic>MALDI-MS</topic><topic>Mass Spectrometry - methods</topic><topic>Miscellaneous</topic><topic>Molecular Sequence Data</topic><topic>Proteins</topic><topic>Proteins - analysis</topic><topic>Proteins - genetics</topic><topic>Silver staining</topic><topic>Staining and Labeling - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hellman, Jukka</creatorcontrib><collection>AGRIS</collection><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>MEDLINE - Academic</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hellman, Jukka</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Polyacrylamide lamination enables mass spectrometry compatible staining and in-gel digestion of proteins separated by agarose IEF</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2007-10-01</date><risdate>2007</risdate><volume>7</volume><issue>19</issue><spage>3441</spage><epage>3444</epage><pages>3441-3444</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>Agarose IEF enables the separation of large proteins and protein complexes. A complication of agarose gels attached onto polyester support is the lack of sensitive protein staining methods compatible with protein analysis and identification protocols. In this study, agarose IEF gels were used to separate the proteins, followed by layering the agarose with polyacrylamide. The formed laminate gels were seamless and durable and they were readily detached from the polyester. The gels were amenable to MS compatible staining. The sensitivity obtained with the acidic silver staining method was 20-50 ng/band of myoglobin. Laminated agarose was a suitable matrix for in-gel digestion based generation of tryptic peptides for MALDI-MS.</abstract><cop>Weinheim</cop><pub>Wiley-VCH Verlag</pub><pmid>17722206</pmid><doi>10.1002/pmic.200700299</doi><tpages>4</tpages></addata></record> |
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| ispartof | Proteomics (Weinheim), 2007-10, Vol.7 (19), p.3441-3444 |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Acrylic Resins - chemistry Agarose IEF Amino Acid Sequence Analytical, structural and metabolic biochemistry Biological and medical sciences Electrophoresis, Agar Gel - methods Fundamental and applied biological sciences. Psychology GelBond In-gel digestion Isoelectric Focusing - methods MALDI-MS Mass Spectrometry - methods Miscellaneous Molecular Sequence Data Proteins Proteins - analysis Proteins - genetics Silver staining Staining and Labeling - methods |
| title | Polyacrylamide lamination enables mass spectrometry compatible staining and in-gel digestion of proteins separated by agarose IEF |
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