Cell Signaling Nuclear Receptor Pathway Tools Nuclear Receptor Analysis Luciferase Vectors Product pGL4.36[luc2P/MMTV/Hygro] Vector pFN26A (BIND) hRluc-neo Flexi® Vector pBIND-ERα Vector pBIND-GR Vector pGL4.35[luc2P/9XGAL4UAS/Hygro] Vector Size Cat.# Price (Fr) 20 µg E1360 612.00 20 µg E1380 612.00 20 µg E1390 612.00 20 µg E1581 612.00 20 µg E1370 612.00 GloResponse™ 9XGAL4UAS-luc2P HEK293 Cell Line 2 vials E8530 6721.00 For Research Use Only. Not for Use in Diagnostic Procedures. Description: Nuclear receptor analysis can be performed with traditional means by using a minimal promoter vector with nuclear receptor response elements upstream. Alternatively, you can use viral elements like the mouse mammary tumor virus long terminal repeat promoter to judge androgen or glucocorticoid responses (e.g., pGL4.36). In many cases, study with these methods requires use of a cell line with the appropriate endogenous nuclear receptors, meaning you may need different cell lines for each nuclear receptor study. A method using the principles of the yeast two-hybrid system was adapted for nuclear receptor work. The nuclear receptor ligand binding domain is fused to the GAL4 DNA binding domain and transfected with a firefly luciferase vector containing repeats of the GAL4 upstream activation sequence upstream of a minimal promoter. The ligand binding domain is responsible for ligand binding, homo- or heterodimerization and interactions with co-activator or co-repressors. The one-hybrid method allows you work with any cell line and nuclear receptor you desire. Features: • Robust: GAL4-based system removes background signals from endogenous receptors. • More Sensitive: Optimized 9X Gal4 gives improved responses, better signal:noise ratio. • Adaptable: Combination Renilla/Neomycin marker allows normalization with Dual-Luciferase® Assay or selectable markers for generating stable cell lines, all with one vector. • Consistent: Compare or profile all nuclear receptors with a single experimental system. • Faster Results: Destabilized and optimized luc2P luciferase gene allows greater sensitivity and shorter induction times. Storage Conditions: Store at –20°C. Protocol pGL4.36[luc2P/MMTV/Hygro] Vector Protocol pFN26A (BIND) hRluc-neo Flexi Vector Protocol pBIND-ERα Vector Protocol pBIND-GR Vector Protocol pGL4.35[luc2P/9XGAL4UAS/Hygro] Vector Protocol GloResponse™ 9XGAL4UAS-luc2P HEK293 Cell Line Technical Bulletin Part# 9PIE136 9PIE138 9PIE139 9PIE158 9PIE137 TB552 pmirGLO Dual-Luciferase miRNA Target Expression Vector Product pmirGLO Dual-Luciferase miRNA Target Expression Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The pmirGLO Vector is designed to quantitatively evaluate microRNA (miRNA) activity by the insertion of miRNA target sites downstream or 3´ of the firefly luciferase gene (luc2). Firefly luciferase is the primary reporter gene; reduced firefly luciferase expression indicates the binding of endogenous or introduced miRNAs to the cloned miRNA target sequence. This vector is based on Promega dual-luciferase technology, with firefly luciferase (luc2) used as the primary reporter to monitor mRNA regulation and Renilla luciferase (hRluc-neo) acting as a control reporter for normalization and selection. Features: • Measure the function of miRNA and miRNA binding sites. • luc2 luciferase gene provides highest expression. • Combination Renilla/Neomycin marker allows for normalization with Dual-Luciferase® Assay or selectable markers for generating stable cell lines, all with one vector. • Moderate-strength PGK promoter provides sensitive analysis not possible with strong promoters. Storage Conditions: Store at –20°C. Protocol Part# pmirGLO Dual-Luciferase miRNA Target Expression Vector Protocol 9PIE133 Size Cat.# Price (Fr) 20 µg E1330 612.00 15 For complete and up-to-date product information visit: www.promega.com/catalog 273 Reporter Assays and Transfection Pagina 276

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