Source:http://linkedlifedata.com/resource/pubmed/id/15450817
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
2
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| pubmed:dateCreated |
2004-9-28
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| pubmed:abstractText |
Heparanase plays an important role in the degradation of the extracellular matrix. It is implicated in inflammation, tumor angiogenesis and metastasis. We have developed two high-throughput methods for measuring heparanase activity and screening potential inhibitors. The first method involves coating fibroblast growth factor (FGF) on microtiter plates and capturing fluorescein isothiocyanate (FITC)-labeled heparin sulfate (HS), which is used as a substrate for heparanase digestion. Labeled HS fragments are released into the medium and quantitated by fluorescence intensity measurement. We have implemented this assay method into a Zeiss uHTS system and screened compound libraries for heparanase inhibitors. The second method involves labeling HS with biotin followed by FITC to generate a dual-labeled HS. The labeled material is bound to streptavidin-coated plates and used as a substrate for heparanase digestion. Both methods are sensitive and easily applicable to robotic systems. In addition, we have labeled both HS and biotin-HS with Eu-chelate, a fluorophore that exhibits long decay fluorescence. Assays using Eu-labeled HS and Eu-labeled biotin-HS have been developed and show higher sensitivity than those using FITC-labeled material. Furthermore, assays using Eu-chelate HS (or biotin-HS) should eliminate the interference of fluorescence compounds.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Anti-Inflammatory Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Antineoplastic Agents,
http://linkedlifedata.com/resource/pubmed/chemical/Biotin,
http://linkedlifedata.com/resource/pubmed/chemical/Fibroblast Growth Factors,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescein-5-isothiocyanate,
http://linkedlifedata.com/resource/pubmed/chemical/Glucuronidase,
http://linkedlifedata.com/resource/pubmed/chemical/Heparitin Sulfate,
http://linkedlifedata.com/resource/pubmed/chemical/Tritium,
http://linkedlifedata.com/resource/pubmed/chemical/heparanase
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| pubmed:status |
MEDLINE
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| pubmed:month |
Oct
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| pubmed:issn |
0003-2697
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
15
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| pubmed:volume |
333
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
389-98
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| pubmed:dateRevised |
2005-11-17
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| pubmed:meshHeading |
pubmed-meshheading:15450817-Animals,
pubmed-meshheading:15450817-Anti-Inflammatory Agents,
pubmed-meshheading:15450817-Antineoplastic Agents,
pubmed-meshheading:15450817-Biotin,
pubmed-meshheading:15450817-CHO Cells,
pubmed-meshheading:15450817-Cattle,
pubmed-meshheading:15450817-Cells, Cultured,
pubmed-meshheading:15450817-Cloning, Molecular,
pubmed-meshheading:15450817-Cricetinae,
pubmed-meshheading:15450817-Drug Evaluation, Preclinical,
pubmed-meshheading:15450817-Escherichia coli,
pubmed-meshheading:15450817-Fibroblast Growth Factors,
pubmed-meshheading:15450817-Fluorescein-5-isothiocyanate,
pubmed-meshheading:15450817-Gene Expression,
pubmed-meshheading:15450817-Glucuronidase,
pubmed-meshheading:15450817-Heparitin Sulfate,
pubmed-meshheading:15450817-Humans,
pubmed-meshheading:15450817-Kidney,
pubmed-meshheading:15450817-Staining and Labeling,
pubmed-meshheading:15450817-Tritium
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| pubmed:year |
2004
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| pubmed:articleTitle |
High-throughput methods for measuring heparanase activity and screening potential antimetastatic and anti-inflammatory agents.
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| pubmed:affiliation |
Department of Discovery Technologies, Hoffmann-La Roche Inc., 340 Kingsland Street, Nutley, NJ 07110, USA. kuo-sen.huang@roche.com
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| pubmed:publicationType |
Journal Article
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