Science Catalog 2012 Life Cell Signaling Chroma-Luc™ Vectors Product pCBR-Basic Vector pCBR-Control Vector pCBG68-Basic Vector pCBG68-Control Vector pCBG99-Basic Vector pCBG99-Control Vector For Research Use Only. Not for Use in Diagnostic Procedures. Description: The Chroma-Glo™ Luciferase Assay System and the Chroma-Luc™ Vectors can be used to generate red and green (dual-color) luminescence from a single sample upon addition of a single reagent. The Chroma-Luc™ Vectors consist of 6 plasmids containing synthetic versions of a red or one of two green click beetle luciferase genes; CBRluc contains a red-emitting luciferase gene, while CBG68luc and CBG99luc contain greenemitting luciferase genes. Filtered measurement of the dual-color luminescence produced by the Chroma-Luc™ luciferases permits each reporter to be measured independently and virtually simultaneously. Besides their different luminescence colors, the three Chroma-Luc™ genes differ as follows: CBG99luc and CBRluc possess 99% DNA and 98% protein homology and are the ideal choice for use when working with transient expression assays; CBG68luc and CBRluc possess 68.9% DNA homology while retaining a high degree of protein homology (>98%) and thus are the preferred pair for use with stable expression assays. Each of these genes is provided either in a Basic Vector configuration containing a multiple cloning site (MCS) or a Control Vector containing an SV40 promoter and enhancer. The Chroma-Glo™ Assay has a homogeneous format that generates luminescence with >30-minute signal half-lives for each of the Chroma-Luc™ Luciferases, thereby enabling the processing of many plates without prior sample preparation. Two reporter gene measurements can be efficiently and reproducibly determined from each well in a typical high-throughput screen. Features: • Two Reporter Signals by Single Substrate Addition: Increase your accuracy and precision through normalization, or use both reporters to multiplex experimental measurements. Use filters to spectrally separate the luminescent signals. • Ideal Control or Multiplexed Reporter System: Use the high-homology red and green luciferases to minimize potential RNA and protein effects on reporter expression. • Flexible: Use the Basic Vectors for cloning regulatory elements of interest, or use the Control Vectors as an internal control. • High Expression with Minimal Anomalous Transcription Behavior: Use the synthetic gene design to obtain results easily and reliably. Storage Conditions: Store at –20°C. Protocol Chroma-Luc™ Reporter Vectors Technical Manual Part# TM059 Size Cat.# Price (Fr) 20 µg E1411 646.00 20 µg E1421 646.00 20 µg E1431 646.00 20 µg E1441 646.00 20 µg E1451 646.00 20 µg E1461 646.00 SalI BamHI poly(A) signalHpaI SV40 late XbaI CBG68luc or CBRluc or CBG99luc B. Ampr f1 ori ori SalI BamHI Enhancer SV40 poly(A) signalHpaI SV40 late XbaI CBG68luc or CBRluc or CBG99luc The Chroma-Luc™-Basic and -Control Vectors. These vectors contain CBRluc or CBG68luc or CBG99luc; Ampr, a gene conferring ampicillin resistance in E. coli; ori, origin of plasmid replication in E. coli. Arrows within the Chroma-Luc™ and Ampr genes indicate the direction of functionality. * MluI should not be used in the vector configuration containing CBG99luc, as this gene also contains the MluI site. Chroma-Luc™-Control Vector Configurations Synthetic poly(A) signal/transcriptional pause site (for background reduction) KpnI SacI MluI* NheI SmaI XhoI BgIII SV40 Promoter HindIII NcoI A. Ampr f1 ori ori Chroma-Luc™-Basic Vector Configurations Synthetic poly(A) signal/transcriptional pause site (for background reduction) KpnI SacI MluI* NheI SmaI XhoI BgIII HindIII NcoI 274 For complete and up-to-date product information visit: www.promega.com/catalog 4220MA06_3A Pagina 277
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