Human NFkB Secreted Alkaline Phosphatase Reporter SEAP - (SEAPorter™) Stable Reporter Cell Line

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Ligand Activation: Human NFkB Secreted Alkaline Phosphatase SEAP - (SEAPorter™) Stable Reporter Cell Line [NBP2-26260] - Evaluation of the functional activity of the NF-kB/SEAP Stable Reporter Cell Line. ...read more

Product Details

Summary
Applications LA
Host
HEK293

Order Details

Human NFkB Secreted Alkaline Phosphatase Reporter SEAP - (SEAPorter™) Stable Reporter Cell Line Summary

Description
Placental alkaline phosphatase is one of the most stable isoenzyme, only existing in the placenta of higher primates. These characteristics make placental alkaline phosphatase suitable to use as a reporter gene for the analysis of promoter activity and gene expression in cell culture and animal serum. The natural form of placental alkaline phosphatase (PLAP) is membrane anchored. The recombinant form of placental alkaline phosphatase (secreted alkaline phosphatase, SEAP) is used for reporter gene function. SEAP is created by inserting a translational terminator after amino acid 489 (Berger, et al., Gene 66 (1): 10 (1988). This mutation converts the membrane-bound PLAP protein into the secreted protein.

As a major transcription factor, NF-kB plays a key role in regulating genes responsible for the innate and adaptive immune responses. In unstimulated cells, the NF-kB dimers are held in the cytoplasm by IkBs that masks the nuclear localization signals of NF-kB. Upon cell stimulation, which leads to IkB degradation, NF-kB quickly translocates to the nucleus and activates various genes that have DNA-binding sites for NF-kB.

The NF-kB/SEAP stable HEK 293 cell line is designed to measure NF-kB activation using SEAP protein secreted to the culture media as a read-out with our SEAPorter™ Assay kit (NBP2-25285). The NF-kB/SEAP stable cells are not only useful in helping with the identification of pro or anti-inflammatory substances, but also can help to assay for proteasome activity since the activation of NF-kB results in the degradation of IkB through the proteasome-dependant pathway.

Contents: 3~4 x 10^6 cells
Biosafety Level: 2
Immunogen
The NF-kB/SEAP is a reporter gene assay for the detection of NF-kB activation and contains a cell line modified for this purpose. This cell line is derived from HEK 293 (Human Embryonal Kidney) cells. It is stably transfected with the SEAP (secreted alkaline phosphatase) reporter gene under the transcriptional control of an NF-kB response element. The recombinant cell line is provided frozen.
Target Species
Human
Specificity
NF-kB/SEAP
Selection Agent
G418
RCL Type
SEAP - (SEAPorter™)
Reporter Gene
Secreted alkaline phosphatase (SEAP)
Host
HEK293

Applications/Dilutions

Agonist
Readout System
Publications
Read Publications using
NBP2-26260 in the following applications:

Packaging, Storage & Formulations

Storage
Store in gas phase of liquid nitrogen.
Reconstitution Instructions
Complete Growth Medium: DMEM with 4.5 g/L glucose + 10% FBS + 4 mM L-glutamine + 1 mM sodium pyruvate + 100 units/ml penicillin + 0.1 mg/ml streptomycin + 0.5 mg/ml G418 (Geneticin).

Details for NFkB Secreted Alkaline Phosphatase Reporter

Type
Cell

Notes

Assume all cultures are hazardous since they may harbor latent viruses or other organisms that are uncharacterized. The following safety precautions should be observed.
- Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
- No eating, drinking or smoking while handling cells.
- Wash hands after handling cultures and before leaving the lab.
- Decontaminate work surface with disinfectant or 70% ethanol before and after working with cells.
- All waste should be considered hazardous.
- Dispose of all liquid waste after each experiment and treat with bleach.

Alternate Names for NFkB Secreted Alkaline Phosphatase Reporter

  • SEAP assay

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Reporter Cell Lines are guaranteed for 1 year from date of receipt.

Publications for NFkB Secreted Alkaline Phosphatase Reporter Reporter Cell Line (NBP2-26260)(5)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 2 applications: ELISA, In vitro.


Filter By Application
ELISA
(1)
In vitro
(1)
All Applications
Filter By Species
Human
(3)
All Species
Showing Publications 1 - 5 of 5.
Publications using NBP2-26260 Applications Species
Zurcher C, Sauter KS, Schweizer M. Pestiviral Erns blocks TLR-3-dependent IFN synthesis by LL37 complexed RNA. Veterinary Microbiology. 2014 Oct 13 [PMID: 25457366]
Signorino G, Mohammadi N, Patane F et al. Role of TLR13 in innate immune recognition of group B streptococci. Infect. Immun. 2014 Sep 15 [PMID: 25225249] (In vitro, Human)

Details:
Human NFkB Secreted Alkaline Phosphatase SEAP - (SEAPorter™) Stable Reporter Cell Line was transiently transfected with full-length mouse TLR13 and stimulated with heat-killed bacteria and bacterial RNA or with phorbol myristate acetate /PMA followed by quantification of NFkB Secreted Alkaline Phosphatase using ELISA kit for the latter.
In vitro Human
Rockman S, Dyson A, Koernig S et al. Evaluation of the bioactivity of influenza vaccine strains in vitro suggests that the introduction of new strains in the 2010 Southern Hemisphere Trivalent Influenza Vaccine is associated with adverse events. Vaccine. 2014 Jun 24 [PMID: 24928062] (ELISA, Human)

Details:
Figs 5,6: The HEK293 NF-kB/SEAP reporter cells were stimulated with various influenza strains from the 2005-2012 flu season. Note: Recombinant TNF-alpha, endotoxin, and TLR ligands poly I:C and R848 were used as positive stimulation controls for activating the reporter cell line. The authors also did some additional characterization of the cell line, and the cell line tested positive for the endogenous presence of TLR3, TLR8, MDA5,and RIG-I but not TLR7 by RT-PCR.
ELISA Human
Laguette N, Bregnard C, Hue P et al. Premature activation of the SLX4 complex by Vpr promotes G2/M arrest and escape from innate immune sensing. Cell. 2014 Jan 16 [PMID: 24412650] (Human)

Details:
NF-kB SEAP activity, Figs S6D, E, G.
Human
Andre CM, Greenwood JM, Walker EG et al. Anti-inflammatory procyanidins and triterpenes in 109 apple varieties. J Agric Food Chem. 2012 Oct 24 [PMID: 23013475]

Details:
Product cited, NF-kB/SEAPorter HEK293 Cell Line (IML-101): Fig 3, Table 1, S2. Readout assay: The study evaluated the potential of apple varieties to reduce inflammation. Phenolic and triterpenes from apple flesh were analyzed for their ability to inhibit

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