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使用 PRESTO-Tango Assay 在 GPCR-宽范围范围内对 β-Arrestin2 进行配制筛查的平行审讯
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Parallel Interrogation of β-Arrestin2 Recruitment for Ligand Screening on a GPCR-Wide Scale using PRESTO-Tango Assay
DOI:

09:03 min

March 10, 2020

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Chapters

  • 00:05Introduction
  • 00:43Primary Screening: Poly-L-Lysine Plate Preparation
  • 01:25Primary Screening: Cell Culture and Plate Seeding
  • 02:22Primary Screening: DNA Plate Preparation and Transfections
  • 03:35Primary Screening: Cell Stimulation and Luminescence Reading
  • 04:26Secondary Screening: Cell Seeding and Transfections
  • 05:59Secondary Screening: Half Log Dose-Curve Drug Plate Preparation and Luminescence Reading
  • 07:16Results: Representative Primary Screening Hit Identification and Secondary Screening Dose-Curve Responses
  • 08:23Conclusion

Summary

Automatic Translation

鉴于全球化学品资源是有吸引力的可药物靶点,因此GPCR配体筛查对于确定铅化合物和脱化研究是必不可少的。为了这些努力,我们描述了PRESTO-Tango,一个开源资源平台,用于使用基于TEV的记者分析,同时分析在大约300个全球资源中的瞬态β-逮捕2招募。

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