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| KINASE ASSAY DEVELOPMENT, SCREENING AND PROFILING |
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| Protein kinases are involved in regulation of multiple cellular processes by catalyzing the phosphorylation of target substrates. As a result, the dysregulation of kinases is often associated with varieties of disease disorders, e.g. neoplasias. Selective protein kinase inhibitors therefore offer an opportunity to target these diseases in the active field of drug discovery. |
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| HDB offers comprehensive services from kinase assay development, HTS to kinase selectivity profiling. All the assays developed by HDB are compatible for HTS. The quality of the service, quick turnaround time, and competitive prices have earned HDB high marks from its existing clients. |
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A: ATP Km determination for kinase A using ADP Quest format.
B: Establishment of initial velocity for the determination of kinase A ATP Km.
C: Kinase A titration using Kinase Glo Plus format.
D: Kinase profiling-IC50 determination using Kinase Glo Plus.
PKAcb inhibitor titration was performed in 384-well plate in a total volume of 10μl.Curve fitting was performed using GraphPad Prism 5 software. |
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| Kinase Assay Development |
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Biochemical assays |
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Formats: Kinase Glo Plus, IMAP, ADP-Quest, Z-Lyte etc. |
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Assay development includes enzyme kinetic analysis (i.e. Km determination for both ATP and substrate), enzyme titration and |
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assay validation. |
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Assay development is applicable for a broad range of kinases involved in MAPK pathway, cell cycle, NF-kB pathway, |
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AKT/PKB pathway, JAK/STAT, membrane receptor, etc. |
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Cell based assays |
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ELISA is routinely used for cell based assay development |
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Transient transfection and generation of stable cell lines are used for overexpression of genes of interest |
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| Screening Serveices |
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| Kinase Selectivity Profiling |
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The diversified panels of 20, 30, 50 or more kinases for profiling service |
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Single compound test and library screening |
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Single dose measurement and IC50 determination |
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Profiling format: Kinase Glo Plus, IMAP or Z'Lyte |
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