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- W1967157579 abstract "We describe here detailed protocols to design, optimize and validate in vitro phosphatase assays that we have utilized to conduct high-throughput screens for inhibitors of dual-specificity phosphatases: CDC25B, mitogen-activated protein kinase phosphatase (MKP)-1 and MKP-3. We provide details of the critical steps that are needed to effectively miniaturize the assay into a 384-well, high-throughput format that is both reproducible and cost effective. In vitro phosphatase assays that are optimized according to these protocols should satisfy the assay performance criteria required for a robust high-throughput assay with Z-factors >0.5, and with low intra-plate, inter-plate and day-to-day variability (CV <20%). Assuming the availability of sufficient active phosphatase enzyme and access to appropriate liquid handling automation and detection instruments, a single investigator should be able to develop a 384-well format high-throughput assay in a period of 3-4 weeks." @default.
- W1967157579 created "2016-06-24" @default.
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- W1967157579 date "2007-05-01" @default.
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- W1967157579 title "Development and optimization of high-throughput in vitro protein phosphatase screening assays" @default.
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- W1967157579 doi "https://doi.org/10.1038/nprot.2007.155" @default.
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