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TOPFlash Lentiviral Dual-Reporter System
$2,400.00
Discover the next generation of reporter assays with our advanced TOPFlash Luciferase Reporter, uniquely designed to simplify your workflow and deliver unparalleled reliability. Unlike traditional systems that require multiple constructs and complicate normalization, our innovative all-in-one design integrates both the Firefly Luciferase and GLuc internal control in a single, efficient construct. This groundbreaking approach eliminates the variability in genome integration and transduction efficiency, ensuring more consistent and trustworthy data. Experience the superior signal strength and convenience only LipExoGen’s proprietary enhancer and integrated system can provide. Upgrade your assays, reduce complexity, and achieve more reliable results with our cutting-edge TOPFlash Luciferase Reporter.
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Available Options:
Specifications
Details
The TOPFlash Lentiviral Dual-Reporter System is an innovative upgrade on the widely used TOPFlash assay, combining an all-in-one luminescent or fluorescent dual-reporter construct, and the broad transduction advantages provided by high-titer VSV-G pseudotyped lentivirus. The reporter system is supplied as a set of two vials of high-titer, prepackaged lentiviral particles including one vial containing the experimental (Wnt-responsive) reporter system, and one vial containing a negative control (TCF/LEF mutant or “FOP”) reporter system. Each vial contains 380 ul of highly purified and concentrated lentivirus in serum-free RPMI medium, which is suitable for transduction of even difficult-to-transfect cell types, such as primary or non-dividing cells.
This product is offered in either a luminescent or fluorescent format to suit your specific application or instrument requirements. The luminescent set utilizes Firefly Luciferase (FLuc) and Gaussia Luciferase (GLuc) reporters, whereas the fluorescence set utilizes a combination of EGFP and mCherry reporters.
The lentiviral particles in vial 1 are packaged with the experimental construct, which contains 6X tandem repeats of TCF/LEF response elements (Wnt response elements, or “WRE”) coupled to a minimal TATA promoter and upstream proprietary enhancer element to drive Wnt-responsive expression of the Firefly Luciferase reporter. Within the same construct, a separate expression cassette utilizes the human EF1a promoter to drive constitutive expression of Gaussia Luciferase and a blasticidin resistance gene (BSD), which are separated by a P2A element and translated as two separate proteins.
Unlike traditional TOPFlash dual-reporter systems (which require two separate constructs and utilize Renilla Luciferase as the internal control), the Gaussia Luciferase in our all-in-one construct functions as a true internal control, eliminating the need for co-transfection or co-/sequential transductions with conventional plasmid or lentiviral based systems, respectively. Gaussia Luciferase utilizes the same substrate as Renilla Luciferase (coelenterazine) and is detected at the same wavelength (481 nm for Renilla, 485 nm for Gaussia), making this reporter system convenient for integrating into existing Dual-Luciferase workflows. By consolidating the Wnt-responsive reporter and internal control into a single construct, our TOPFlash Dual-Reporter System accounts for both variable transduction efficiency and site of genomic integration, which cannot be achieved using two separate viruses. Like other commercially available TOPFlash reporter systems, the internal control can also be used to reflect differences in cell viability resulting from experimental treatments. Our proprietary Gaussia Luciferase variant is retained intracellularly, and can therefore be detected in cell lysates in the same manner as Renilla luciferase in typical TOPFlash assays.
The lentiviral particles in vial 2 are packaged with a mutant TCF/LEF reporter (“FOP”) construct, which is identical to the experimental construct except that the multimerized response elements are mutated such that Wnt responsive Firefly Luciferase expression is abolished. This allows the user to control for false positives by defining any baseline expression of Firefly Luciferase in the absence of Wnt activity.
In the fluorescent variant, TCF/LEF response elements (or their mutated counterpart) drive expression of EGFP, whereas constitutively expressed mCherry serves as the internal control.
Both versions of this product contain blasticidin resistance gene to facilitate generation of stable reporter cell lines.
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| TRS-0001 | |
| High-titer Lentiviral Particles, third generation, VSV-G pseudotyped | |
| Serum-free RPMI-1640, frozen solution | |
| One set containing 2 vials of lentiviral particles (one experimental and one negative control) | |
| 380 μl/vial | |
| 2×10^6 TU/vial, depending on the variant | |
| 3×10^8 VP/ml | |
| -80oC |
*Based on infectivity on HEK293FT cells transduction units (TU).
Construct Layouts
Advantages
- All-in-one Reporter System Consolidated FLuc + GLuc internal control within the same construct provides faster and more efficient lentiviral transduction, accelerating the time to stable reporter cell lines, eliminating variability in reporter expression associated with two separate constructs, such as differences in transduction efficiency, copy number, and site of genomic integration.
- Versatile Readouts Luminescent (FLuc + GLuc) or fluorescent (EGFP + mCherry) dual readouts provide unprecedented flexibility.
- Compatible with Mainstream GLuc is easily detected like RLuc in commercially available alternative dual-reporter assays, using the same substrate and wavelength detection.
- Innovative Technology Utilizes our novel reporter technology with upstream enhancer and 6X TCF/LEF response element to boost signal intensity.
- License-free & Affordable Generate your own cell lines with ease, freeing you from constraints of expensive fixed commercial reporter cell lines. The EF1α promoter-driven GLuc and BSD resistance provides long-term stable expression in cell lines.
- Unmatched customization Various reporters and selection markers to choose from, perfectly tailored to your specific research needs.
Custom Orders
If you require a modification to one of our products, such as a change in reporter or other vector component, please contact us. Examples of potential customization options are shown in the table below and can be custom-made upon request. Please note that not all options may be feasible due to size limitations and this will depend on the specific product. We also welcome requests for custom products like the one shown on this page.
Additional Custom Service Options
- Send us your cells and we can establish a stable reporter cell line for you using this product. Learn more.
- ORF cDNA plasmids featured in the product figures are available upon request.
- Ultra-high concentration virus can be provided upon request in your choice of medium and volume (i.e. for in vivo applications).
Lentiviral Transduction Protocol
MSDS
Frequently Asked Questions
Frequently Asked Questions
Can I purchase the plasmid for this product?
TF reporter products such as this one are not available in plasmid format. They are only available as high-titer lentiviral particles. Other gene products on our site, such as ORF cDNA and shRNA products are available in either format.