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时间分辨Förster共振能量转移测定法，用于测量人细胞中的内源性磷酸化STAT蛋白

JoVE Journal
Bioquímica

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JoVE Journal Bioquímica

Time-resolved Förster Resonance Energy Transfer Assays for Measurement of Endogenous Phosphorylated STAT Proteins in Human Cells

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时间分辨Förster共振能量转移测定法，用于测量人细胞中的内源性磷酸化STAT蛋白

DOI:

10.3791/62915-v

•

07:12 min

•

September 09, 2021

•

Jaime Padros, Geneviève Chatel, Mireille Caron

¹BioAuxilium Research

Capítulos

00:05Introduction
01:02Stimulator or Inhibitor Titration Using the Two-Plate Assay Protocol with Adherent Cells
04:33Results: Two-Plate and One-Plate Detection of STAT Proteins in Adherent and Suspension Cell Lines, and Intraplate Variability Study
06:40Conclusion

Summary

Tadução automática

描述了基于时间分辨Förster共振能量转移细胞的测定方案，用于以384孔格式对细胞裂解物中内源性磷酸化信号转导和转录激活子（STAT）1/3/4/5/6蛋白的有效定量进行简单，特异性，灵敏和稳健的定量。

Tags

Keywords: Time-resolved FRET TR-FRET STAT Proteins JAK/STAT Signaling Immunoassay Cell Culture Cell Stimulation Cell Lysis Phosphatase Inhibitors High-throughput Screening Pharmacological Characterization

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