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Thermo Scientific™ pCMV-Red Firefly Luc Vector for Luciferase Assays
Description
The Thermo Scientific™ pMCS-Red Firefly Luc Vector is a multiple cloning site plasmid designed to accept a promoter sequence for study of gene regulation using the intracellular red firefly luciferase reporter.
The Red Firefly Luc Vectors contain a mutant form of the firefly luciferase gene that has a red-shifted emission spectrum. The pMCS vector contains a multiple cloning site for cloning a promoter to study its regulatory potential. The pCMV and pTK vectors have the luciferase gene under the CMV (Cytomegalovirus) promoter and Herpes Simplex Virus (HSV) thymidine kinase (TK) promoter, respectively. These constitutive expression vectors can be used as normalization controls to account for experimental variation in combination with other reporters.
Highlights:
- Intracellular Red Firefly Luciferase gene, optimized for high expression in mammalian systems
- Multiple cloning site (MCS) provides versatility for transfer of regulatory elements from one plasmid to another
- Transcription termination site (Ter), Lac operator (Lac O1), and transcriptional pause site (TPS) used to minimize background by reducing transcriptional read-through
- Both puromycin (Pur) and ampicillin (Amp) markers for drug selection in mammalian and bacterial cells, respectively
- High-copy pUC bacterial DNA replication origin
- Two control vectors available with strong (CMV) or weak (TK) constitutive promoters for co-transfection and normalization
Specifications
Specifications
| Description | pCMV-Red Firefly Luc Vector |
| Type | Luc Vector |
| Cloning Method | Restriction Enzyme/MCS |
| Delivery Type | Transfection |
| Quantity | 10 μg |
| Content And Storage | Store in original container protected from direct sunlight in a dry, cool and well-ventilated area, between the following temperatures: -70 to -20°C. |
| Reporter Gene | Red Firefly Luciferase |
| Selection Agent (Eukaryotic) | Puromycin |
| Product Line | Pierce |
| For Use With (Application) | Reporter Assays |
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Frequently Asked Questions (FAQs)
There are three vectors available for each expressed luciferase; pMCS, pCMV, and pTK. We recommend using pMCS to measure the activity of the reporter, and pCMV, and pTK as positive control vectors. For high level constitutive expression of Luciferase, we recommend using the CMV (Cytomegalovirus) promoter vector and for low level constitutive expression of luciferase, we recommend using the pTK (thymidine kinase) promoter vector.
Gaussia, Gaussia-Dura, and Cypridina Luciferase are stable in the culture media for greater than 24 hours. Generally, after transfection of CMV driven constructs, we see ever increasing secreted signal between 24 and 72 hours.
The earliest detectable signal will depend on the strength of the promoter. In general, with a strong promoter (such as CMV), significant signal over background can be seen in as little as 20 minutes. However, with a weaker inducible promoter, significant signal over background may take 1-2 hours after induction.
Our qIP Luciferase assay kits have been discontinued but we do carry epitope-tagged (HA or c-Myc) and Tluc (TurboLuc luciferase)-tagged mammalian expression vectors, and qIP protein interaction assay reagents for these assay kits.
Safety and Handling
For Research Use Only. Not for use in diagnostic procedures.
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