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Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms
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Neuroscience
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JoVE Journal Neuroscience
Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms

Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms

DOI:
10.3791/56795-v

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06:40 min

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January 25, 2018

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, , , , , ,
1Laboratory of Molecular Neurobiology, Institute of Cellular and Organismic Biology,Academia Sinica

Chapters

  • 00:05Title
  • 01:17Measurement of Luciferase Reporter Signals, which Correspond to γ-Secretase Cleavage of APP-C99 (CG cells) or NΔE (NG cells)
  • 03:42Quantification of γ-Secretase Activity
  • 04:40Results: Treatments with CL-387, 785 can Differentially Promote the Processing of APP-C99 by γ-Secretase without Affecting Notch Cleavage
  • 06:05Conclusion

Summary

Automatic Translation

Automatic Translation

We have successfully generated two substrate-specific γ-secretase assays. Both cell-based assays presented here are designed to quantify γ-secretase enzymatic activities via the output of firefly luciferase reporters.

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Keywords: Gamma-secretaseAmyloid Precursor ProteinNotch CleavageCell-based Luciferase Reporter AssayAlzheimer’s DiseaseGamma-secretase ModulatorsSubstrate SelectivityNeuroblastomaNG CellsCG CellsTetracyclineDMSOCL-387785DAPTErbB1/ErbB2 InhibitorsLuciferase AssayLuminescence Microplate Reader

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