||The Notch signaling pathway controls cell fate decisions in vertebrate and invertebrate tissues. Notch signaling is triggered through the binding of a transmembrane ligand, such as Delta like ligand 4 (DLL4) or jagged 1 (JAG1) to a 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, RBPJ, Suppressor of Hairless, Lag-1) and the coactivator, Mastermind, to turn on transcription of Notch responsive genes.
||The Notch Pathway Reporter kit is designed for monitoring the activity of Notch signaling pathway in the cultured cells. The kit contains the transfection-ready expression vector for Notch1 with the entire extracellular domain deleted (Notch1DE). Inside the cells, the Notch1DE can be cleaved by -secretase and active Notch1 NICD is released to nucleus. The kit also contains a CSL (CBF1/RBP-J) 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-J) 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 constitutivelyexpressing 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.
||Screen for activators or inhibitors of the Notch signaling pathway.Study effects of RNAi or gene overexpression on the activity of the Notch pathway.
||Stable at least 12 months from date of receipt, when stored as directed (-20°C)
||Reporter (Component A): CSL (CBF1/RBP-Jκ) luciferase reporter vector + constitutively expressing Renilla luciferase vector; 500 µl (60 ng DNA/ µl); -20°C Negative Control Reporter (Component B): Non-inducible luciferase vector + constitutively expressing Renilla luciferase vector; 500 µl (60 ng DNA/ µl); -20°C Notch1∆E (Component C): Expression vector for intracellular domain of Notch1; 250 µl (100 ng DNA/µl); -20°C Negative Control Expression vector (Component D): Empty expression vector without Notch1; 250 µl (100 ng DNA/µl); -20°CThese vectors are designed for use in transient transfections. They are NOT SUITABLE for transformation and amplification in bacteria.