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293/hTLR4-HA

293/hTLR4-HA Unit size Cat. code Docs Qty Price
HEK 293 cells stably transfected with the HA-tagged human TLR4 gene
3-7 x 10e6 cells
293-htlr4ha
+-
$873.00

HEK-293 cells stably transfected with human HA-tagged TLR4a gene

293/hTLR4-HA cells are designed for studying the stimulation of human TLR4 (hTLR4).

293/hTLR4-HA cells were generated by stable transfection of the HEK293 cell line with the hTLR4a (long isoform, canonical sequence) gene fused at the 3’ end to the influenza hemaglutinine (HA) tag. This tag is the epitope of a very efficient and specific monoclonal antibody.
The use of human HA-tagged TLR4a provides a simple and convenient method to detect the expression of the TLR4a gene by Western blot using an anti-HA Tag antibody. Addtion of the HA tag has no deleterious effect on the expression and function of the TLR4 gene.

Notes:
- HEK293 cells express endogeneous levels of TLR1, TLR3, TLR5, TLR6 and NOD1.
- The control cell line for 293/hTLR4-HA cells is 293/null.

 

1. Poltorak A. et al., 1998. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science, 282(5396):2085-8
2. Shimazu R. et al., 1999. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med, 189(11):1777-82
3. Horng T. GM. Barton, and R. Medzhitov, 2001. TIRAP: an adapter molecule in the Toll signaling pathway. Nat Immunol, 2(9):835-41
4. Fitzgerald KA. et al., 2003. LPS-TLR4 Signaling to IRF-3/7 and NF-{kappa}B Involves the Toll Adapters TRAM and TRIF. J Exp Med. 198(7):1043-1055.

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Specifications

Antibiotic resistance: blasticidin

Growth medium: DMEM, 4.5 g/l glucose, 2-4 mM L-glutamine, 10% (v/v) fetal bovine serum, 50 U/ml penicillin, 50 μg/ml streptomycin, 100 μg/ml Normocin™, 2 mM L-glutamine

Quality control:

  • TLR4-HA expression is validated by Western blot.
  • TLR4 activity is validated upon stimulation with LPS 24 hours after co-transfection of 293/hTLR4-HA cells with a human CD14/MD-2 expression plasmid and an NF-kB-inducible SEAP (secreted embryonic alkaline phosphatase) reporter plasmid.
  • These cells are guaranteed mycoplasma-free.
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Contents

Shipped on dry ice

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Description

TLR4, the first human TLR identified, is the receptor for Gram-negative lipopolysaccharide (LPS). The TLR4 gene was shown to be mutated in C3H/HeJ and C57BL/10ScCr mice, both of which are low responders to LPS [1]. However, TLR4 alone is not sufficient to confer LPS responsiveness. TLR4 requires MD-2, a secreted molecule, to functionally interact with LPS [2]. Furthermore, a third protein, called CD14, was shown to participate in LPS signaling, leading to NF-κB translocation. This signaling is mediated through several adaptor proteins: MyD88 TIRAP/Mal [3] , TRIF/TICAM1 and TRAM/TICAM2 [4].

 

1. Poltorak A. et al., 1998. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science, 282(5396):2085-8
2. Shimazu R. et al., 1999. MD-2, a molecule that confers lipopolysaccharide responsiveness on Toll-like receptor 4. J Exp Med, 189(11):1777-82
3. Horng T. GM. Barton, and R. Medzhitov, 2001. TIRAP: an adapter molecule in the Toll signaling pathway. Nat Immunol, 2(9):835-41
4. Fitzgerald KA. et al., 2003. LPS-TLR4 Signaling to IRF-3/7 and NF-{kappa}B Involves the Toll Adapters TRAM and TRIF. J Exp Med. 198(7):1043-1055.

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