Human TLR2 Stable Cell Line

Product Discontinued

Human TLR2 Stable Cell Line Summary

• Description: The TLR2 stable cell line can be used for TLR2 flow cytometric calibration and detection control as well as TLR2-dependent functional assays. TLR2 expression in these cells has been validated by Western blotting (fig. 1) and flow cytometry (fig. 2). Functional activity of this cell line has been validated using the NF-kB/SEAPorter™ Assay Kit (NBP2-25286, fig. 3).
• Immunogen: The TLR2 stable cell line is a stably transfected cell line which expresses full-length human Toll-like receptor 2 (TLR2) with an N-terminal HA tag.
• Target Species: Human
• Specificity: TLR2
• Selection Agent: Blasticidin
• Growth Properties: Adherent Morphology: Epithelial
• RCL Type: Stable Cell Line
• Host: HEK293
• Gene: TLR2

Applications/Dilutions

• Agonist:
  • MALP-2, TLR6 and TLR2 ligand NBP2-26219
• Readout System:
  • NFkB Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) NBP2-25286
  • Cell Surface Staining Flow Cytometry Kit NBP2-26247

Packaging, Storage & Formulations

• Storage: Store in gas phase of liquid nitrogen.
• Reconstitution Instructions: Comlete 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 TLR2 stable line.
- Wash hands after handling the TLR2 stable 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 Stable 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 (NBP2-26266)(1)

Publication using NBP2-26266

• Mukherjee S, Biswas R, Biswas T. Alternative TLRs are stimulated by bacterial ligand to induce TLR2-unresponsive colon cell response. Cell Signal. 2013-08-01 [PMID: 23665374] (IP/WB)
  
  Details:
  TLR Stable cell lines cited in Fig 2C for IP/WB:1. TLR1 Stable Cell Line (IML-201)2. TLR2 Stable Cell Line (IML-202)3. TLR4 Stable Cell Line (IML-204)Note: The cell lines were exposed to porin and IP'd with a TLR1, TLR2, or TLR4 antibody, respectively, fo

FAQs for TLR2 Reporter Cell Line (NBP2-26266). (Showing 1 - 1 of 1 FAQ).

1. The customer stimulated the products TLR4 Stable Cell Line (NBP2-26268) and TLR2 Stable Cell Line (NBP2-26266) with their ligand, and he detected the expression of Defencin by semi-quantitative PCR. Defencin is the response factor downstream of TLR signaling. However the expression level is same to normal HEK cell. When he also used HaCat cell, the expression level of Defencin increased commensurately. He suggests the HaCat cell line seems to be better response than these products. Do you know why this is? Also, TLR2 exists in complex with TLR1 or TLR6 in native condition. TLR4 exists in complex with MD2 and CD14. He noticed whether the TLR in these cell line are not over-expressd, because these cell lines does not over-express their complex element together. He would like to make sure that the expression level of TLR in IMGENEX TLR cell line is higher than native cell line. Additionally he wants to know whether TLR can be detected by using TLR antibody in western blotting generally. Tag antibody can be not used for such analysis, because Tag antibody can not detect native endogenous TLR. Coud you please tell us your suggestion?
   • Functional activity of this cell line requires co-transfection of MD-2 and CD14. With regards to readout assays, we have evaluated both the NBP2-26268 (TLR4) and NBP2-26266 (TLR2) with the SEAP reporter assays as shown on the respective data sheets. We don't perform semi-quantitative PCR and I am sorry we don't have any information regarding the ligand the customer used nor a Defencin PCR quantitative PCR readout assay. Hence, it would be difficult to comment further on the customer's results. We would suggest that the customer use the techniques and assays shown on the data sheet as positive controls for cell line function. This includes the SEAP reporter assays which require co-transfection of the NF-kB SEAP reporter, and for the TLR4 cell it also requires co-transfection of MD2 and CD14. This also includes using established TLR4 (LPS) and TLR2 (PAM3CSK4). Once the researcher establishes the positive control system, they can move to testing their ligand with the SEAP readout assay and then to their Defencin readout assay. However, it must be recognized that the suitability of the cell lines for a researcher's model system needs to be empirically determined and optimized by the researcher. Both the NBP2-26266 and NBP2-26268 cell lines are selected for function as assayed by a SEAP reporter assay (see data on data sheets). Overexpression results are shown by flow cytometry using the appropriate TLR antibodies with respect to the Vector control cell line. Please refer to the data sheets for these results. For WB, we use an HA Tag antibody. The TLRs were not recognized by WB using the TLR antibodies we evaluated. We would suggest using a Tag antibody if the researcher wishes to detect the TLR constructs expressed by the cell lines. We are not studying endogenous TLRs in the cell lines, the focus is on the functional aspects of the TLR genes stably transfected into the cell lines. If the customer wishes to use TLR antibodies to assess the TLRs, then flow cytometry assays should be performed as shown on the data sheets. On another note, HEK293 do not express functional TLR2 or TLR4. I am not sure what ligand the researcher is using but activation of normal HEK293 cells might be occurring through other pathways.

Bioinformatics

• Gene Symbol: TLR2