Notch Reporter BioAssay™ Kit (cAMP/PKA Pathway)

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.

Kit Components:
171935A: Reporter, CSL (CBF1/RBP-Jkappa), 1x500ul
171935B: Negative Control, 1x500ul
171935C: Notch1∆E, 1x250ul
171935D: Negative Control Expression vector, 1x250ul

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.