The pCBR-Control, pCBG68-Control and pCBG99-Control Vectors, where the reporter gene is expressed from the SV40 enhancer and promoter, are used as internal controls in a dual-reporter system. Cells transfected with Chroma-Luc™ Vectors and used with the Chroma-Glo™ Luciferase Assay System reagent can be used to generate red and green (dual-color) luminescence from a single sample. Use the Basic Chroma-Luc™ Vectors as an experimental reporter for the Control Vectors.
By having two reporter signals from adding a single substrate, you can increase your accuracy and precision through normalization, or use both reporters to multiplex experimental measurements. Use filters to spectrally separate the luminescent signals. The synthetic gene design of the Chroma-Luc™ Vectors offers high expression with minimal anomalous transcription behavior. The Chroma-Luc™ genes offer ideal control or multiplexed reporter system because the high-homology red and green luciferases minimize potential RNA and protein effects on reporter expression.
Note: Use of this product requires a luminometer fitted with filters. See the Chroma-Luc™ Reporter Vectors Technical Manual #TM059 for information on recommended filter sets.
About the Synthetic Click Beetle Luciferase Genes
The Chroma-Luc™ Vectors consist of six 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 green-emitting 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.
Chroma-Luc™ Reporter Vectors Technical Manual
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