Microarrayed compound screening (microARCS) to identify activators and inhibitors of AMP-activated protein kinase
| Autor: | Usha Warrior, Ernst U. Frevert, William J. Chiou, David J. Burns, David A. Egan, Leo W. Barrett, Barbara L. Cool, Lemma Kifle, Paul L. Richardson, Steven N. Anderson, James L. Kofron |
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| Rok vydání: | 2004 |
| Předmět: |
0301 basic medicine
Streptavidin Time Factors High-throughput screening AMP-Activated Protein Kinases Protein Serine-Threonine Kinases 01 natural sciences Biochemistry Sensitivity and Specificity Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Adenosine Triphosphate Multienzyme Complexes Animals Protein kinase A Oligonucleotide Array Sequence Analysis chemistry.chemical_classification Miniaturization AMPK 0104 chemical sciences Rats 010404 medicinal & biomolecular chemistry 030104 developmental biology Enzyme chemistry Biotinylation Molecular Medicine Agarose Adenosine triphosphate Biotechnology |
| Zdroj: | Journal of biomolecular screening. 9(2) |
| ISSN: | 1087-0571 |
| Popis: | A novel and innovative high-throughput screening assay was developed to identify both activators and inhibitors of AMP-activated protein kinase (AMPK) using microarrayed compound screening (microARCS) technology. Test compounds were arrayed at a density of 8640 on a polystyrene sheet, and the enzyme and peptide substrate were introduced into the assay by incorporating them into an agarose gel followed by placement of the gels onto the compound sheet. Adenosine triphosphate (ATP) was delivered via a membrane, and the phosphorylated biotinylated substrate was captured onto a streptavidin affinity membrane (SAM trade mark ). For detection, the SAM trade mark was removed, washed, and imaged on a phosphor screen overnight. A library of more than 700,000 compounds was screened using this format to identify novel activators and inhibitors of AMPK. |
| Databáze: | OpenAIRE |
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