背景介绍 The 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 the transcription of Notch-responsive genes. 产品介绍 Notch Pathway Reporter kit is designed for monitoring the activity of the Notch signaling pathway in cultured cells. The kit contains a transfection-ready expression vector for Mouse NOTCH1 that has a deletion of the entire extracellular domain and transmembrane region (NOTCH1 NICD). Inside the cells, the NOTCH1 NICD is constitutively localized into the nucleus without needing to be cleaved by γ-secretase. The kit also contains CSL (CBF1/RBP-Jκ) 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-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 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. This kit contains the expression vector for Mouse Notch1 NICD, which does not require γ-secretase processing to become active. We also offer the Mouse Notch1DE expression vector (BPS Bioscience #60509), as well as a Human Notch1DE expression vector (BPS Bioscience #79503), which are dependent on γ-secretase activity to become active.