Human TLR2 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line

Product Discontinued

Human TLR2 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line Summary

• Description: The TLR2 reporter line can be used for TLR2-dependent functional assays as well as screening of TLR2 agonists or antagonists. The TLR2 reporter cell line has been validated by flow cytometry (fig. 3) and ligand dose response assay (Fig. 2 and Fig. 3).
  
  Contents: 3~4 x 10^6 cells
  Biosafety Level: 2
• Immunogen: The TLR2 reporter cell line is a stably co-transfected cell line which expresses full-length human Toll-like receptor 2 (TLR2) and the secreted alkaline phosphatase (SEAP) reporter gene under the transcriptional control of an NF-kB response element.
• Target Species: Human
• Specificity: TLR2/NF-kB/SEAP
• Selection Agent: Blasticidin and G418.
• Growth Properties: Adherent Morphology: Epithelial
• RCL Type: NF-kB/SEAP - (SEAPorter™)
• Reporter Gene: Secreted alkaline phosphatase (SEAP)
• Host: HEK293
• Gene: TLR2

Applications/Dilutions

• Dilutions:
  • Flow Cytometry
  • Ligand Activation
• Agonist:
  • Pam3CSK4, TLR1 and TLR2 Ligand NBP2-25297
  • Zymosan, TLR2 Ligand NBP2-26233
  • Histone NBP2-26236
  • MALP-2, TLR6 and TLR2 ligand NBP2-26219
• Readout System:
  • Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) NBP2-25285

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 + 10 ug/ml blasticidin + 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 the reporter cell line.
- Wash hands after handling the reporter cell line 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 Human TLR2 NF-kB/SEAP - (SEAPorter™) Stable Reporter Cell Line

• CD282 antigen
• CD282
• TIL4CD282
• TLR2
• toll/interleukin 1 receptor-like 4
• Toll/interleukin-1 receptor-like protein 4
• toll-like receptor 2

Background

Toll-like receptors are type I transmembrane signaling receptors which are critical for the innate host defense to pathogens. Toll-like receptor 2 (TLR2), known as CD282, has been identified as a receptor that is central to the innate immune system's response to lipoproteins of Gram-negative bacteria and Gram-positive bacteria, as well as a receptor for peptidoglycan and lipoteichoic acid and other bacterial cell membrane products. The TL2.1 antibody is useful for blocking studies. It has been reported to block TLR2 agonist-induced cellular activation.

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 TLR2 Reporter Cell Line (NBP2-26274)(4)

Publications using NBP2-26274

• Yamaguchi M, Hirose Y, Takemura M, et al Streptococcus pneumoniae Evades Host Cell Phagocytosis and Limits Host Mortality Through Its Cell Wall Anchoring Protein PfbA. Front Cell Infect Microbiol. 2019-08-20 [PMID: 31482074] (Bacteria)
  
  Details:
  Mice were infected with S. pneumoniae.
• Crowe JS, Roberts KJ, Carlton TM et al. Preclinical Development of a Novel, Orally-Administered Anti-Tumour Necrosis Factor Domain Antibody for the Treatment of Inflammatory Bowel Disease. Sci Rep. 2018-03-21 [PMID: 29563546] (Human)
• Chan M, Hayashi T, Mathewson RD et al. Identification of substituted pyrimido[5,4-b]indoles as selective Toll-like receptor 4 ligands. J Med Chem. 2013-06-13 [PMID: 23656327]
• Greene CM, Carroll TP, Smith SG et al. TLR-induced inflammation in cystic fibrosis and non-cystic fibrosis airway epithelial cells. J Immunol. 2005-02-01 [PMID: 15661927]
  
  Details:
  Functional Assays: 1.TLR2/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-102). 2. TLR4/MD-2/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-104MD2). 3. TLR4/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-104).4.TLR7/NF-kB/SEAP Stably Transfected HEK 293 Cells (IML-107). 5. LPS from E. Coli, TLR4 Ligand (IMG-2204). 6. SEAPorter Assay Kit (10055K).

FAQs for TLR2 Reporter Cell Line (NBP2-26274). (Showing 1 - 3 of 3 FAQs).

1. Which promoter drives to express TLR on TLR expression vector into the TLR/NF-kB/SEAP Stable Reporter Cell Line. What is the vector which is delivered into this cell line?
   • The TLR vectors are derived from a CMV promoter-based mammalian expression vector. Hence the promoter for the TLR genes is CMV. However, the exact sequence of the vector is proprietary.
2. The protocol describes that cells settle down in 1-3 days; however, the cells took longer to grow. Why is this?
   • The time to settle down can vary and can take longer than 1-3 days depending on a number of variables, including cell culture conditions, cell recovery from shipment/storage, etc. One - 3 days is an approximate time. It would be impossible to predict exactly how long it would take the cells to settle down in a given investigators hands and conditions. Remember the cells have travelled around the world and outside of their usual liquid nitrogen storage, and then are brought out of storage into culture. They are exposed to many variables throughout that process, they make take up to 5-7 days to settle down.
3. We would like to know whether this cell line expresses other TLRs endogenously. Could you let us know if there is any information?
   • About NBP2-26274 (Human TLR2 NF-kB/SEAP - (SEAPorter) Stable Reporter Cell Line), it was derived from the stably co-transfected cell line (HEK 293) which expresses full-length human Toll-like receptor 2 (TLR2) and the secreted alkaline phosphatase (SEAP) reporter gene under the transcriptional control of an NF-kB response element. This product was first developed by Imgenex which is now a part of Novus. I don't think Imgenex has tested the expression of other TLRs, but I anticipate that the reason HEK 293 cell line was chosen was the minimal expression of TLR2 in vivo or the endogenous protein would affect the reporter assay.

Bioinformatics

• Gene Symbol: TLR2