Monitoring notch activation in cultured mammalian cells: Luciferase complementation imaging assays

Ma Xenia G. Ilagan, Raphael Kopan

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Notch activation and cleavage releases the Notch intracellular domain (NICD), which translocates to the nucleus, where it associates with its DNA-binding partner CSL to recruit the coactivator MAML and additional cofactors to ultimately activate target gene expression. Taking advantage of the specific interaction between NICD and these factors, we have developed a luciferase complementation imaging (LCI)-based reporter system to quantitatively monitor Notch activation in real time in live cells. In this chapter, we describe the use of Notch LCI reporters for measuring protein interactions and performing detailed kinetic analyses of receptor activation and its responses to various stimuli.

Original languageEnglish
Title of host publicationNotch Signaling
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages155-168
Number of pages14
ISBN (Print)9781493911387
DOIs
StatePublished - 2014

Publication series

NameMethods in Molecular Biology
Volume1187
ISSN (Print)1064-3745

Keywords

  • Bioluminescence
  • LCI
  • Luciferase complementation imaging
  • Molecular imaging
  • Notch
  • PCA
  • Protein complementation assay
  • Protein-protein interaction
  • Split luciferase

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    Ilagan, M. X. G., & Kopan, R. (2014). Monitoring notch activation in cultured mammalian cells: Luciferase complementation imaging assays. In Notch Signaling: Methods and Protocols (pp. 155-168). (Methods in Molecular Biology; Vol. 1187). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-1139-4_12