The Notch Pathway Reporter BioAssay™ kit is designed for monitoring the activity of the Notch signaling pathway in cultured cells. The kit contains a transfection-ready expression vector for NOTCH1 that has a deletion of the entire extracellular domain (Notch1deltaE). Inside the cells, the NOTCH1 DE can be cleaved by upsilon-secretase and active NOTCH1 NICD is released into the nucleus. The kit also contains CSL (CBF1/RBP-Jk) luciferase reporter vector, which is a Notch pathway-responsive reporter. This reporter contains the firefly luciferase gene under the control of multimerized CSL responsive elements upstream of a minimal promoter. The CSL (CBF1/RBP-Jk) 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 non-inducible 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.
Applications:
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.
Note: These vectors are useful for transient transfection. They are not suitable for transformation and amplification in bacteria.
Storage and Stability:Store all components at -20°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
