Human TLR9 Stable Cell Line

Product Discontinued

Human TLR9 Stable Cell Line Summary

• Description: The TLR9 stable cell line can be used for TLR9 flow cytometric calibration and detection control as well as TLR9-dependent functional assays. TLR9 expression in this cell line has been validated by flow cytometry (fig. 1). Functional activity of this cell line has been validated using the NF-kB/SEAPorter™ Assay Kit (NBP2-25286, fig. 2).
  
  Contents: 3~4 x 10^6 cells
  Biosafety Level: 2
• Immunogen: The TLR9 stable cell line is a stably transfected HEK 293 line which expresses full-length human Toll-like receptor 9 (TLR9) with no epitope tag.
• Target Species: Human
• Specificity: TLR9
• Selection Agent: Blasticidin
• Growth Properties: Adherent Morphology: Epithelial
• RCL Type: Stable Cell Line
• Host: HEK293
• Gene: TLR9

Applications/Dilutions

• Agonist:
  • CpG oligodeoxynucleotides with negative control, TLR9 ligand NBP2-26232
• Readout System:
  • NFkB Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) NBP2-25286
  • Intracellular Staining Kit NBP2-26248

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.

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 stable line.
- Wash hands after handling the stable line and before leaving the lab.
- Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.
- All waste should be considered hazardous.
- Dispose of all liquid waste after each experiment and treat with bleach.

Alternate Names for Human TLR9 Stable Cell Line

• CD289 antigen
• CD289
• TLR9
• toll-like receptor 9

Background

Toll-like receptors (TLRs) are evolutionarily conserved pattern-recognition molecules resembling the toll proteins that mediate antimicrobial responses in Drosophila. These proteins recognize different microbial products during infection and serve as an important link between the innate and adaptive immune responses. TLR9 forms a subfamily along with TLR7 and TLR8 that recognize viral RNA and CpG DNA sequences and are localized in intracellular acidic compartments such as the phagolysosome. Unlike other TLRs which act through adaptor molecules such as TOLLIP, TIRAP, TRIF, and MyD88 to activate various kinases and transcription factors to respond to potential infection, TLR9 is strictly dependent on MyD88.

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 TLR9 Reporter Cell Line (NBP2-26272)(2)

Publications using NBP2-26272

• Daley D, Zambirinis CP, Seifert L et al. gammadelta T Cells Support Pancreatic Oncogenesis by Restraining alphabeta T Cell Activation. Cell 2016-08-25 [PMID: 27569912] (FLOW, Mouse, Human)
• Feng D, Sangster-Guity N, Stone R et al. Differential requirement of histone acetylase and deacetylase activities for IRF5-mediated proinflammatory cytokine expression. J Immunol. 2010-11-15 [PMID: 20935208]

FAQs for TLR9 Reporter Cell Line (NBP2-26272). (Showing 1 - 1 of 1 FAQs).

1. Could you tell us expression site of TLR9 of the cell line? In general, TLR9 expresses in endosome, but we would like to know if the cell line was forcibly expressed TLR9 in cell membrane.
   • About "expression site of TLR9", we don't know where the gene was integrated in the genome of HEK293 cells unfortunately.  I anticipate that when the cell line clone was selected, Imgenex (the original company which produced this stable cell line. Novus acquired it last year) there must have many colonies that were also picked after stably transfecting with the TLR9 plasmid. After many testing, this clone was expanded and provided to the customer as NBP2-2627 (Human TLR9 Stable Cell Line) as it behaved just like the parental cell line, I believed. Due to the proprietary information, the plasmid that was used in the stable transfection experiments and whether the integration site was determined have never been disclosed to us. When transfecting plasmids in the mammalian cells, regardless transient and stable transfection, the scientists hope to obtain a fractions of cells that could harbor the plasmids of interest (just the limited numbers) that could reach to the nuclei. The nuclei are the place for the transfected genes to be expressed (of course the same place where the gene can be integrated) Since the TLR9 gene in the plasmid was not tagged with any epitope and it is a full-length gene PDF, the expressed TLR9 protein should behave just like any human proteins: goes to the location(s) where they should reside. More specially it should behave like the endogenous TLR9 protein. For answering your question about "... TLR9 expresses in endosome, but we would like to know if the cell line was forcibly expressed TLR9 in cell membrane", I would like to point out that the expression of the TLR9 gene in the nuclei will lead to the TLR9 hnRNA synthesis in the organelle, followed by the transportation/processing of hnRNA to cytoplasm and then protein synthesis. 

Bioinformatics

• Gene Symbol: TLR9