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基于NMR的活性测定,用于测定核苷裂解酶的化合物抑制、IC50值、伪影活性和全细胞活性
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JoVE Journal Chemistry
NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases
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基于NMR的活性测定,用于测定核苷裂解酶的化合物抑制、IC50值、伪影活性和全细胞活性

DOI:
10.3791/59928-v

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10:24 min

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June 30, 2019

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, , , , , , , , , , ,
1Department of Chemistry,Adelphi University, 2Department of Chemistry,Washington University in St. Louis, 3Department of Chemistry,Boston University

Chapters

  • 00:04Title
  • 00:59Initial Test Compound Assay
  • 05:01Detergent Counter Screen Assay
  • 06:19Jump-Dilution Counter Screen Assays
  • 07:56Results: Representative Nuclear Magnetic Resonance (NMR)-Based Assay Results
  • 09:28Conclusion

Summary

Automatic Translation

Automatic Translation

基于NMR的活性测定已经开发出来,以识别和表征两种核苷糖酶的抑制剂。为500μM和250μM的初始化合物测定、确定IC50值的剂量反应测定、洗涤剂计数器屏幕测定、跳跃稀释计数器筛查测定以及大肠杆菌全细胞测定提供了协议。

Tags

NMR-based Activity AssaysEnzyme InhibitionIC50 ValuesArtifact DetectionWhole-cell ActivityNucleoside RibohydrolasesFragment ScreeningTrichomonas VaginalisAnti-trichomonal DrugsReagent VolumesReaction ComponentsSubstrate PreparationTest Compound PreparationReaction Stock SolutionReaction InitiationReaction Quenching

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