NEW PRODUCT ALERT!
SBI is labored as quite a bit as announce the launch of our subsequent interval of pGreenFire signalling pathway reporters! We’ve upgraded these customary lentivectors with a clever design that allows reliable interval of ordinary cell traces and have moreover modified the standard luciferase reporter with crimson firefly luciferase, which opens up the potential of performing a dual-spectral luciferase assay and delivers bigger sensitivity for in vivo choices than typical luciferase.
Overview
Make your personal pGreenFire 2.zero reporter vector or use as a detrimental administration
With the pGreenFire 2.zero mCMV Cloning & Unfavorable Administration Lentivector & Virus (pGF2-mCMV-rFluc-T2A-GFP-mPGK-Puro), likelihood is excessive it’s possible you’ll revenue from our sturdy pGreenFire 2.zero lentivector know-how to create your personal transcriptional response half (TRE) reporter or use pGreenFire 2.zero mCMV as-is as a detrimental administration.
SBI Lentiviral Applied sciences
With the pGreenFire 2.zero mCMV Cloning & Unfavorable Administration Lentivector, XhoI and NheI web websites are positioned upstream of a minimal CMV promoter (mCMV) so likelihood is excessive it’s possible you’ll clone in your personal TREs. Upon activation, the TREs and mCMV promoter collectively drive co-expression of crimson firefly luciferase and GFP so likelihood is excessive it’s possible you’ll quantitatively measure transcriptional put together using every fluorescence and luciferase put together.
GreenFire Fundamentals
For lots of who’re not acquainted with our pGreenFire reporters, every 1.zero and a few.zero share an related core effectivity—transcriptional response components (TREs) are positioned upstream of a minimal CMV promoter (mCMV) and the pGreenFire luciferase-T2A-GFP co-expression cassette. All by way of the absence of transcriptional activation, the mCMV promoter has negligible put together resulting in little- to no- luciferase put together or GFP fluorescence (Resolve 1). Nonetheless, upon activation of the TREs, akin to in response to the addition of an inducer, the TREs plus the mCMV promoter drive expression of every luciferase and GFP in a dose-dependent pattern (Resolve 1). The result is the pliability to quantitatively measure pathway activation using luciferase put together or whereas imaging using GFP.
As with all of our pGreenFire 2.zero lentivectors, the GreenFire cassette now consists of crimson firefly luciferase (rFLuc), a T2A co-expression half, and GFP. The swap to rFLuc opens up the potential of performing a dual-spectral luciferase assay and likewise delivers bigger sensitivity for in vivo choices than typical luciferase.
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