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Time-resolved Förster Resonance Energy Transfer Assays for Measurement of Endogenous Phosphorylated STAT Proteins in Human Cells
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生物化学
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JoVE 杂志 生物化学
Time-resolved Förster Resonance Energy Transfer Assays for Measurement of Endogenous Phosphorylated STAT Proteins in Human Cells

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

DOI:
10.3791/62915-v

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07:12 min

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September 09, 2021

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, ,
1BioAuxilium Research

Chapters

  • 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

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Time-resolved Förster resonance energy transfer cell-based assay protocols are described for the simple, specific, sensitive, and robust quantification of endogenous phosphorylated signal transducer and activator of transcription (STAT) 1/3/4/5/6 proteins in cell lysates in a 384-well format.

Tags

Keywords: Time-resolved FRETTR-FRETSTAT ProteinsJAK/STAT SignalingImmunoassayCell CultureCell StimulationCell LysisPhosphatase InhibitorsHigh-throughput ScreeningPharmacological Characterization

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