Promega, N1471, Coincidence Reporter Vectors

Name: Coincidence Reporter Vectors
Brand: Promega
SKU: N1471

Vectors That Express Both Firefly and NanoLuc® Luciferases from the Same Transcript, Firefly and NanoLuc® luciferase have dissimilar compound interference profiles for better identifying more false-positives versus individually, Using two different transcriptional reporters reduces false hit rates and increases the identification of true biological hits, luc2, and, NlucP, provide a bright reporter combination compatible with low-copy-number and plate scale-up, and provide greater signal-to-backg

Catalog Number: N1471

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Category

Engineered Cell Lines

TagsLuciferase Assays Genetic Reporter Vectors and Cell Lines

Description

pNLCoI reporter vectors express the firefly luciferase and the destabilized NanoLuc® luciferase reporters on the same mRNA transcript. The two reporters are separated by a short P2A peptide, which causes ribosome skipping and the generation of two soluble, unfused forms of luciferase with distinct inhibitor profiles.
Two Reporter Genes, One Transcript
Luciferase-based reporter-gene assays remain a useful and powerful method of high-throughput compound screening. However, false hits that result from direct interaction of compounds with the luciferase reporter can result in unnecessary follow-up efforts. The pNLCoI Vectors comprise a second-generation coincidence reporter vector system that allow expression of both firefly luciferase ( luc2 ) and NanoLuc® Luciferase fused to a PEST destabilization domain ( NlucP ) from the same mRNA transcript. The stoichiometric expression of both luciferases is achieved by use of the P2A sequence from porcine teschovirus-1, which promotes a ribosomal skip and expression of the two unfused enzymes with distinct compound interaction profiles. When used in high-throughput compound screening, false hits caused by direct interaction with one or the other luciferases can be distinguished from true hits that show a similar response for both, reducing workload associated with follow-up screens.
The pNLCoI Vectors are designed for use with the Nano-Glo® Dual-Luciferase® Reporter (NanoDLR™) Assay System, which sequentially detects firefly and NanoLuc® Luciferase in activity in the same sample with a convenient “add-read-add-read” homogeneous format. Both NanoDLR™ reagents provide stable glow-type luminescence signals with half-lives of approximately two hours for automated batch processing of samples and amenable to assays or high-throughput screens in 96-, 384- or 1,536-well plate formats. Potent inhibition of firefly luciferase coupled with the high-intensity luminescence of NanoLuc® luciferase maximizes sensitivity for detection of both reporters.
pNLCoI1[ luc2 -P2A- NlucP /Hygro] Vector GenBank® Accession Number KM359771 and vector sequence text file .
pNLCoI2[ luc2 -P2A- NlucP /minP/Hygro] Vector GenBank® Accession Number KM359772 and vector sequence text file .
pNLCoI3[ luc2 -P2A- NlucP /CMV/Hygro] Vector GenBank® Accession Number KM359773 and vector sequence text file .
pNLCol4[ luc2 -P2A- NlucP /PGK/Hygro] Vector GenBank® Accession Number KM376434 and vector sequence text file .

Specifications

{"type": "table", "rows": [{"name": "pNLCoI2[ luc2 -P2A- NlucP /minP/Hygro] Vector", "conditions": {"Part #": "N147A", "Size": "1 × 20μg"}}]}

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