293/TLR4-MD2-CD14

HEK293 clones expressing TLR4-MD2-CD14

293/TLR4-MD2-CD14 cells were obtained by stable co-transfection of HEK293 cells with the pUNO-TLR4, which expresses the human or murine TLR4 gene, and the pDUO2-MD2-CD14 plasmid which expresses the human or murine MD2 and CD14 genes.

The control cell line of 293/TLR4-MD2-CD14 cells is 293/null.

Figures

293/TLR, NOD and control clones were transfected transiently with pNiFty-SEAP and stimulated with TLR and NOD ligands. After 16 hour stimulation, NF-κB-induced SEAP activity was assessed using QUANTI-Blue™, a SEAP detection medium.

Specifications

Antibiotic resistance: blasticidin and hygromycin B

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™

Guaranteed mycoplasma-free

These products are covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 100 μl Blasticidin (10 mg/ml)
• 100 μl Hygromycin B Gold (ultra-pure hygromycin B; 100 mg/ml)
• 1 ml Normocin™ (50 mg/ml)

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

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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