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

Product Discontinued

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

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence
  • Ligand Activation
• Application Notes: Use in ICC/IF reported in scientific literature (PMID:33201697)
• Agonist:
  • Flagellin, TLR5 Ligand NBP2-25289
• Readout System:
  • Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) NBP2-25285

Reactivity Notes

Human reactivity reported in scientific literature (PMID:33201697)

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 Array

• 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 Array

• 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)(7)

Publications using NBP2-26260

• Jackson PA, Schares HAM, Jones KFM et al. Synthesis of Guaianolide Analogues with a Tunable alpha-Methylene-gamma-lactam Electrophile and Correlating Bioactivity with Thiol Reactivity Nat Commun 2020-11-13 [PMID: 33201697] (ICC/IF, Human)
• MohanKumar, K;Namachivayam, K;Song, T;Jake Cha, B;Slate, A;Hendrickson, JE;Pan, H;Wickline, SA;Oh, JY;Patel, RP;He, L;Torres, BA;Maheshwari, A; A murine neonatal model of necrotizing enterocolitis caused by anemia and red blood cell transfusions Nat Commun 2019-08-02 [PMID: 31375667]
• Zurcher C, Sauter KS, Schweizer M. Pestiviral Erns blocks TLR-3-dependent IFN synthesis by LL37 complexed RNA. Veterinary Microbiology. 2014-10-13 [PMID: 25457366]
• Signorino G, Mohammadi N, Patane F et al. Role of TLR13 in innate immune recognition of group B streptococci. Infect. Immun. 2014-09-15 [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.
• 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. 2014-06-24 [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.
• 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. 2014-01-16 [PMID: 24412650] (Human)
  
  Details:
  AP-1-Luciferase activity, Figs 6D, F, G
• Andre CM, Greenwood JM, Walker EG et al. Anti-inflammatory procyanidins and triterpenes in 109 apple varieties. J Agric Food Chem. 2012-10-24 [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