pNiFty2-IFA-SEAP
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Cat.code:
pnf2-ifasp
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ABOUT
Inducible SEAP reporter plasmid- mIFNα promoter - ZeocinR
pNiFty2-IFA plasmid is composed of two key elements:
- the mouse interferon alpha (mIFN-α) 4 minimal promoter and
- a SEAP (Secreted alkaline phosphatase) reporter gene.
Interferons are key modulators of the immune response. Their pleiotropic activities are mediated by the induction of many IFN‑stimulated genes (ISGs). To help study the transcriptional regulation and signal transduction of type I IFNs, InvivoGen provides several reporter systems, called pNiFty2, based on the inducible expression of the secreted embryonic alkaline phophatase (SEAP) gene. The SEAP gene is cloned under the control of three different promoters that are activated by various transcription factors, such as IRF3, IRF5, IRF7 and NF‑κB.
pNiFty2‑IFA‑SEAP features the mouse IFN-α promoter. Transcription of mIFN-α4 is mediated by a virus responsive element (VRE-A4) located in the promoter. VRE-A4 contains four cooperating DNA modules that bind to IRF3 adn IRF7. Co‑expression of IFN-α and SEAP with constitutively activated IRF3 (saIRF3) or IRF7 (saIRF7) in HEK293 cells led to a strong increase in SEAP expression.
pNiFty2-IFA plasmids are selectable with Zeocin™ in both E. coli and mammalian cells, and can be used to generate stable clones. For your convenience, InvivoGen also provides the QUANTI-Blue™ reporter detection reagent to measure SEAP activity in the supernatant.
All products are for research use only, and not for human or veterinary use.
SPECIFICATIONS
Specifications
In vitro transfection
Plasmid construct is confirmed by restriction analysis and full-length open reading frame (ORF) sequencing.
CONTENTS
Contents
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Product:pNiFty2-IFA-SEAP
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Cat code:pnf2-ifasp
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Quantity:20 µg
1 ml of Zeocin® (100 mg/ml)
Shipping & Storage
- Shipping method: Room temperature
- -20 °C
- Avoid repeated freeze-thaw cycles
Storage:
Caution:
Details
Minimal promoter
The proximal promoters are shorter than 500 bp and contain transcription factor binding sites. Upon stimulation in 293 cells, their expression level remains undetectable. With the addition of repeated TFBS, the proximal promoters become inducible by the appropriate stimulus and drive the expression of the reporter gene.
mIFNα promoter is the mouse interferon alpha 4 minimal promoter [1]. Transcription of mIFNα4 is mediated by a virus responsive element (VRE-A4) located in the promoter. VRE-A4 contains four cooperating DNA modules that
bind to IRF3 adn IRF7 [2]. Co-expression of IFNα-SEAP with constitutively activated IRF3 (saIRF3) or IRF7 (saIRF7) in HEK293 cells led to a strong increase in SEAP expression.
Reporter Gene
SEAP reporter gene: Secreted alkaline phosphatase (SEAP) is a reporter widely used to study promoter activity or gene expression. SEAP expression can be rapidly and readily measured in supernatants of transfected cells. SEAP levels can be evaluated qualitatively with the naked eye and quantitatively using SEAP detection media, such as HEK-Blue™ Detection system.
References:
1. Braganca J. et al., 1997. Synergism between multiple virus-induced factor-binding elements involved in the differential expression of interferon A genes. J Biol Chem. 272(35):22154-62.
2. Morin P. et al., 2002. Preferential binding sites for interferon regulatory factors 3 and 7 involved in interferon-A gene transcription. J Mol Biol. 316(5):1009-22.
DOCUMENTS
Documents
Technical Data Sheet
Plasmid Sequence
Safety Data Sheet
Certificate of analysis
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