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腺苷酸环化酶的药物引起的过敏性：含量简化和小型化的小分子和siRNA筛选应用

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications

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腺苷酸环化酶的药物引起的过敏性：含量简化和小型化的小分子和siRNA筛选应用

DOI:

10.3791/51218-v

•

09:39 min

•

January 27, 2014

•

Jason M. Conley, Tarsis F. Brust, Ruqiang Xu, Kevin D. Burris, Val J. Watts

¹Department of Medicinal Chemistry and Molecular Pharmacology,Purdue University, ²Quantitative Biology,Eli Lilly and Company

Chapters

00:05Title
01:37Expansion and Cryopreservation of Assay Ready Cells
02:58Plating Assay Ready Cells
04:15Small Molecule Screening and Persistent Agonist Treatment
05:16Stimulation, Quenching, and Estimation of cAMP Accumulation
07:02Results: Development and Testing of a Cell-based 384 Well Sensitization Assay with Receptor Antagonists
09:12Conclusion

Summary

Automatic Translation

持续激活的抑制性G蛋白偶联受体导致腺苷酸环化酶信令的致敏作用。以确定必需的分子途径，无偏差的方法是必要的，然而，这种策略需要一个可扩展的基于细胞的cAMP敏感测定法的发展。本文中，我们描述了一种致敏试验用于小分子和siRNA的筛选。

Tags

Adenylyl Cyclase Sensitization CAMP G Protein-coupled Receptors D2 Dopamine Receptor Opioid Receptor High-throughput Screening SiRNA Assay Miniaturization Cell-based Assay

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