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Human TLR4/MD-2/CD14 SEAP - (SEAPorter™) Stable Reporter Cell Line

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Summary
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    • Catalog Number
      NBP2-26503
    • Availability
      Product Discontinued

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Human TLR4/MD-2/CD14 SEAP - (SEAPorter™) Stable Reporter Cell Line Summary

Description
The TLR4/MD-2/CD14 reporter line can be used for TLR4-dependent functional assays as well as screening of TLR4 agonists or antagonists. The TLR4/MD-2/CD14 reporter cell line has been validated by a ligand dose response assay (Figure 1).

Contents: 3~4 x 10^6 cells
Biosafety Level: 2
Immunogen
The TLR4/MD-2/CD14 reporter cell line is a stably co-transfected cell line which expresses full-length human Toll-like receptor 4 (TLR4), human MD-2, human CD14 and secreted alkaline phosphatase (SEAP) reporter genes under the transcriptional control of an NF-kB response element.
Target Species
Human
Specificity
TLR4/MD-2/CD14/NF-kB/SEAP
Selection Agent
Blasticidin, puromycin, zeocin and G418
Growth Properties
Adherent Morphology: Epithelial
RCL Type
SEAP - (SEAPorter™)
Host
HEK293

Applications/Dilutions

Dilutions
  • Functional
  • In vitro assay
  • Ligand Activation
Application Notes
Use in function and in-vitro reported in scientific literature (PMID 25640528)
Agonist
Readout System
Publications
Read Publications using
NBP2-26503 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 + 10 ug/ml blasticidin + 2 ug/ml puromycin + 0.2 mg/ml zeocin + 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 TLR4/MD-2/CD14 line.
- Wash hands after handling the TLR4/MD-2/CD14 line and before leaving the lab.
- Decontaminate work surface with disinfectant or 70% ethanol before and after working with the TLR4/MD-2/CD14 reporter line.
- All waste should be considered hazardous.
- Dispose of all liquid waste after each experiment and treat with bleach

Alternate Names for Human TLR4/MD-2/CD14 SEAP - (SEAPorter™) Stable Reporter Cell Line

  • TLR4,MD2,CD14

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 TLR4/MD-2/CD14 Reporter Cell Line (NBP2-26503)(6)

We have publications tested in 2 confirmed species: Human, Bacteria.

We have publications tested in 3 applications: Biacore Binding Assay, Func, In vitro.


Filter By Application
Biacore Binding Assay
(1)
Func
(2)
In vitro
(1)
All Applications
Filter By Species
Human
(1)
Bacteria
(1)
All Species
Showing Publications 1 - 6 of 6.
Publications using NBP2-26503 Applications Species
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.
Bacteria
Ryan DA, Degardin M, Alam S et al. Rational Design of Peptide Derivatives for Inhibition of MyD88-Mediated TLR Signaling in Human PBMCs and Epithelial Cells Exposed to F. tularensis. Chem Biol Drug Des. 2017-06-09 [PMID: 28599094] (Biacore Binding Assay, Human) Biacore Binding Assay Human
McKim JM, Wilga PC, Pregenzer JF, Blakemore WR. The common food additive carrageenan is not a ligand for toll-like- receptor 4 (TLR4) in an HEK293-TLR4 reporter cell-line model Food Chem. Toxicol. 2015-01-29 [PMID: 25640528] (Func, In vitro)

Details:
Human TLR4/MD-2/CD14 SEAP - (SEAPorterTM) Stable Reporter Cell Line [HEK293 cells stably transfected with a TLR4/MD-2/CD14/NF-kB/SEAP reporter construct - expresses full-length human TLR4, MD-2, CD14, and secreted alkaline phosphatase/SEAP under the control of an NFkB response element] used for evaluating the ability of different types of Carrageenan/CGN to bind and activate TLR4 signaling. SEAP was measured as an indicator of TLR4 activation with SEAPorterTM assay kit and CGN was found to not to activate the TLR4.
Func, In vitro
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]
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]
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] (Func)

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).
Func

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Product General Protocols

View specific protocols for TLR4/MD-2/CD14 Reporter Cell Line (NBP2-26503): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for TLR4/MD-2/CD14 Reporter Cell Line (NBP2-26503). (Showing 1 - 2 of 2 FAQs).

  1. Do the cell lines carry plasmids and is this why the selection agents are essential to grow the cells. Can the cells expand without losing their plasmids in the absence of selection agents? So what all plasmids are there for what all genes and what is the selection agent for the respective plasmids. Is SEAP also on plasmids?
    • This cell line contains 4 plasmids; TLR4, MD-2, CD14, and NF-kB-SEAP. This is why selection agents are essential when using this cell line. The 4 plasmids are selected using blasticidin, puromycin, zeocin and G418/Geneticin, respectively. SEAP is coupled to NFkB on a plasmid. I would encourage you to use the protocol for this product when growing your cells.
  2. Are there any normalization techniques that can be used in an assay to make the results more reliable and remove the variability of cell numbers?
    • I would recommend that you do not change the media during the whole cell assay. The optimal volume of ligand solution should not exceed 10% of the media volume of each well. In addition, I would recommend that you omit the selection agents during the whole cell assay. The selection agents are required only for cell maintanence.

Blogs on TLR4/MD-2/CD14

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