Notch pathway Reporter Kit
Cat. # 60509

BackgroundThe Notch signaling pathway controls cell fate decisions in vertebrate and
invertebrate tissues. NOTCH signaling is triggered through the binding of a
transmembrane ligand to Notch transmembrane receptor (NOTCH1/
NOTCH2/NOTCH3/NOTCH4) on a neighboring cell. This results in proteolytic
cleavage of the NOTCH receptor, releasing the constitutively active intracellular
domain of NOTCH (NICD). NICD translocates to the nucleus and associates with
transcription factors CSL (CBF1/RBPJκ/Suppressor of Hairless/Lag-1) and
coactivator Mastermind to turn on transcription of Notch responsive genes.

DescriptionNotch Pathway Reporter kit is designed for monitoring the activity of Notch
signaling pathway in the cultured cells. The kit contains the transfection-ready
Notch1 NICD expression vector and CSL (CBF1/RBP-Jκ) luciferase reporter
vector, which is a Notch pathway-responsive reporter. This reporter contains a
firefly luciferase gene under the control of multimerized CSL responsive element
upstream of a minimal promoter. The CSL (CBF1/RBP-Jκ) reporter is premixed
with constitutively expressing Renilla (sea pansy) luciferase vector, which serves
as an internal positive control for transfection efficiency.

The kit also includes a non-inducible firefly luciferase vector premixed with
constitutively-expressing Renilla luciferase vector as a negative control. The noninducible
luciferase vector contains a firefly luciferase gene under the control of a
minimal promoter, but without any additional response elements. The negative
control is critical for determining pathway specific effects and background
luciferase activity.

Application• Monitor Notch signaling pathway activity.
• Screen activators or inhibitors of Notch signaling pathway.
• Study effects of RNAi or gene overexpression on the activity of Notch
pathway.

Components

These vectors are ready for transient transfection. They are NOT MEANT for
transformation and amplification in bacteria.

Reference1. Lu FM et al. (1996) Constitutively active human Notch1 binds to the transcription
factor CBF1 and stimulates transcription through a promoter containing a CBF1-
responsive element. Proc. Natl. Acad. Sci. USA 93(11): 5663-5667.

2. Moellering RE et al. (2009) Direct inhibition of the NOTCH transcription factor
complex. Nature 462(7270):182-188.