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Overvågning af ER / SR-calciumfrigivelse med den målrettede Ca<sup> 2+</sup> Sensor CatchER<sup> +</sup
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JoVE Journal Biology
Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+

Overvågning af ER / SR-calciumfrigivelse med den målrettede Ca<sup> 2+</sup> Sensor CatchER<sup> +</sup

DOI:
10.3791/55822-v

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12:30 min

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May 19, 2017

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, , ,
1Department of Chemistry, Center of Diagnostics and Therapeutics (CDT),Georgia State University

Chapters

  • 00:05Title
  • 01:30Transfection of HEK293 Cells
  • 02:43Transfection of C2C12 Myoblast Cells
  • 04:10Preparation of Slide and Fluorescence Microscope
  • 06:54Imaging Drug-induced Ca2+ Transients and In Situ Kd Calibration
  • 10:14Results: In Situ Kd of CatchER+ in C2C12 Myoblast Cells
  • 11:32Conclusion

Summary

Automatic Translation

Automatic Translation

Vi præsenterer protokoller til anvendelse af vores målrettede genetisk kodede calciumindikator (GECI) CatchER + til overvågning af hurtige calciumtransienter i det endoplasmatiske / sarkoplasmiske retikulum (ER / SR) af HEK293 og skeletmuskulatur C2C12-celler ved hjælp af realtids fluorescensmikroskopi. En protokol til in situ K d måling og kalibrering diskuteres også.

Tags

ER/SR Calcium ReleaseCatchER+HEK293 CellsC2C12 CellsCalcium SignalingCalcium TransientsGECICell TransfectionFluorescence ImagingCalcium-related Diseases

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