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Creating a cell line with an indicator of a functional signaling pathway is useful for deciphering the components in a signaling pathway. These tools are made by inserting multiple repeats of a response element upstream of a minimal promoter (minP). We have designed vectors that report the activity of a variety of pathways using the optimized luc2 firefly luciferase gene in the pGL4 backbone. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P and RapidResponse™ genes are luc2 genes appended with degradation sequences to influence the cellular half-life of firefly luciferase. Proteins encoded by the RapidResponse™ genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity.
These response element vectors also have a hygromycin resistance selectable marker for use either in transient transfection experiments or for selection of a stable cell line. Some of these response element vectors also are available stable cell lines (GloResponse™ Cell Lines) to speed your research. For investigating signaling pathways, we have vectors available that respond to the MAPK/JNK pathway with the pGL4.44[luc2P/AP1 RE/Hygro] Vector, Wnt pathway with the pGL4.49[luc2P/TCF-LEF RE/Hygro] Vector, cAMP/PKA pathway with the pGL4.29[luc2P/CRE/Hygro] Vector, Calcium/Calcineurin pathway with the pGL4.30[luc2P/NFAT-RE/Hygro] Vector, NF-?B pathway with the pGL4.32[luc2P/NF-?B-RE/Hygro] Vector, MAP/ERK pathway with the pGL4.33[luc2P/SRE/Hygro] Vector and RhoA pathway with the pGL4.34[luc2P/SRF-RE/Hygro] Vector.