A method for the covalent capture and screening of diverse small molecules in a microarray format
This protocol describes a robust method for the covalent capture of small molecules with diverse reactive functional groups in microarray format, and outlines a procedure for probing small-molecule microarrays (SMMs) with proteins of interest. A vapor-catalyzed, isocyanate-mediated surface immobiliz...
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| Veröffentlicht in: | Nature protocols 2006-12, Vol.1 (5), p.2344-2352 |
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| creator | Koehler, Angela N Bradner, James E McPherson, Olivia M |
| description | This protocol describes a robust method for the covalent capture of small molecules with diverse reactive functional groups in microarray format, and outlines a procedure for probing small-molecule microarrays (SMMs) with proteins of interest. A vapor-catalyzed, isocyanate-mediated surface immobilization scheme is used to attach bioactive small molecules, natural products and small molecules derived from diversity-oriented synthesis pathways. Additionally, an optimized methodology for screening SMMs with purified proteins and cellular lysates is described. Finally, a suggested model for data analysis that is compatible with commercially available software is provided. These procedures enable a platform capability for discovering novel interactions with potential applications to immunoglobulin profiling, comparative analysis of cellular states and ligand discovery. With the appropriate materials and experimental setup, the printing of SMMs can be completed in 14 hours over 3 days. Screening and data analysis requires 2 days. A detailed timeline is provided. |
| doi_str_mv | 10.1038/nprot.2006.282 |
| format | Article |
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A vapor-catalyzed, isocyanate-mediated surface immobilization scheme is used to attach bioactive small molecules, natural products and small molecules derived from diversity-oriented synthesis pathways. Additionally, an optimized methodology for screening SMMs with purified proteins and cellular lysates is described. Finally, a suggested model for data analysis that is compatible with commercially available software is provided. These procedures enable a platform capability for discovering novel interactions with potential applications to immunoglobulin profiling, comparative analysis of cellular states and ligand discovery. With the appropriate materials and experimental setup, the printing of SMMs can be completed in 14 hours over 3 days. Screening and data analysis requires 2 days. 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Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Koehler, Angela N</au><au>Bradner, James E</au><au>McPherson, Olivia M</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A method for the covalent capture and screening of diverse small molecules in a microarray format</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2006-12</date><risdate>2006</risdate><volume>1</volume><issue>5</issue><spage>2344</spage><epage>2352</epage><pages>2344-2352</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>This protocol describes a robust method for the covalent capture of small molecules with diverse reactive functional groups in microarray format, and outlines a procedure for probing small-molecule microarrays (SMMs) with proteins of interest. A vapor-catalyzed, isocyanate-mediated surface immobilization scheme is used to attach bioactive small molecules, natural products and small molecules derived from diversity-oriented synthesis pathways. Additionally, an optimized methodology for screening SMMs with purified proteins and cellular lysates is described. Finally, a suggested model for data analysis that is compatible with commercially available software is provided. These procedures enable a platform capability for discovering novel interactions with potential applications to immunoglobulin profiling, comparative analysis of cellular states and ligand discovery. With the appropriate materials and experimental setup, the printing of SMMs can be completed in 14 hours over 3 days. Screening and data analysis requires 2 days. A detailed timeline is provided.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>17406478</pmid><doi>10.1038/nprot.2006.282</doi><tpages>9</tpages></addata></record> |
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| source | MEDLINE; Nature Journals Online; SpringerLink Journals - AutoHoldings |
| subjects | Analytical Chemistry Biological Techniques Biomedical and Life Sciences Biomolecules Catalysis Computational Biology/Bioinformatics Data analysis DNA microarrays Identification and classification Isocyanates Life Sciences Ligands Methods Microarrays Natural products Organic Chemistry Polyethylene glycol Protein Array Analysis - methods Protein Binding Proteins Protocol Separation Software |
| title | A method for the covalent capture and screening of diverse small molecules in a microarray format |
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