Cell Signaling Technology

Product Pathways - NF-kB Signaling

Toll-like Receptor 3 (D10F10) Rabbit mAb #6961

Applications Reactivity Sensitivity MW (kDa) Isotype
W H (Mk) Endogenous 115-130 Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Toll-like Receptor 3 (D10F10) Rabbit mAb recognizes endogenous levels of total TLR3 protein. A band is detected at 75 kDa in some cell lines/tissues which is of unknown origin.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Val495 of human TLR3 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from MCF 10A cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-) or SignalSilence® Toll-like Receptor 3 siRNA I #6236 (+), using Toll-like Receptor 3 (D10F10) Rabbit mAb #6961 (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The Toll-like Receptor 3 (D10F10) Rabbit mAb confirms silencing of Toll-like Receptor 3 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with human TLR3 (+), using Toll-like Receptor 3 (D10F10) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-29 cells, untreated or following transfection with pIpC (100 μg/ml; overnight), using Toll-like Receptor 3 (D10F10) Rabbit mAb.


Background

Members of the Toll-like receptor (TLR) family, named for the closely related Toll receptor in Drosophila, play a pivotal role in innate immune responses (1-4). TLRs recognize conserved motifs found in various pathogens and mediate defense responses (5-7). Triggering of the TLR pathway leads to the activation of NF-κB and subsequent regulation of immune and inflammatory genes (4). The TLRs and members of the IL-1 receptor family share a conserved stretch of approximately 200 amino acids known as the Toll/Interleukin-1 receptor (TIR) domain (1). Upon activation, TLRs associate with a number of cytoplasmic adaptor proteins containing TIR domains, including myeloid differentiation factor 88 (MyD88), MyD88-adaptor-like/TIR-associated protein (MAL/TIRAP), Toll-receptor-associated activator of interferon (TRIF), and Toll-receptor-associated molecule (TRAM) (8-10). This association leads to the recruitment and activation of IRAK1 and IRAK4, which form a complex with TRAF6 to activate TAK1 and IKK (8,11-14). Activation of IKK leads to the degradation of IκB, which normally maintains NF-κB in an inactive state by sequestering it in the cytoplasm.

TLR3 functions as a receptor for double-stranded (ds)RNA typically associated with viral infection (4). It was originally shown to be specifically expressed in dendritic cells of the leukocyte family (5). TLR3 has also been found in placenta and lung, and can be induced by LPS in a variety of tissues (4,6). TLR3 is predominantly localized to early endosomes (7,8). Binding of dsRNA, or the analog polyinosine-polycytidylic acid (pIpC), to TLR3 triggers activation of transcription factors NF-κB and IRF3 through the adaptor protein TICAM-1/TRIF (9,10). TRIF associates with members of the TRAF family and with RIP that combine to activate NF-κB and IRF3 (11-13).

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  3. Dunne, A. and O'Neill, L.A. (2003) Sci STKE 2003, re3.
  4. Medzhitov, R. et al. (1997) Nature 388, 394-7.
  5. Schwandner, R. et al. (1999) J Biol Chem 274, 17406-9.
  6. Takeuchi, O. et al. (1999) Immunity 11, 443-51.
  7. Alexopoulou, L. et al. (2001) Nature 413, 732-8.
  8. Zhang, F.X. et al. (1999) J Biol Chem 274, 7611-4.
  9. Horng, T. et al. (2001) Nat Immunol 2, 835-41.
  10. Oshiumi, H. et al. (2003) Nat Immunol 4, 161-7.
  11. Muzio, M. et al. (1997) Science 278, 1612-5.
  12. Wesche, H. et al. (1997) Immunity 7, 837-47.
  13. Suzuki, N. et al. (2002) Nature 416, 750-6.
  14. Irie, T. et al. (2000) FEBS Lett 467, 160-4.
  15. Alexopoulou, L. et al. (2001) Nature 413, 732-8.
  16. Muzio, M. et al. (2000) J Immunol 164, 5998-6004.
  17. Nishimura, M. and Naito, S. (2005) Biol Pharm Bull 28, 886-92.
  18. Matsumoto, M. et al. (2003) J Immunol 171, 3154-62.
  19. Funami, K. et al. (2007) J Immunol 179, 6867-72.
  20. Hoebe, K. et al. (2003) Nat Immunol 4, 1223-9.
  21. Oshiumi, H. et al. (2003) Nat Immunol 4, 161-7.
  22. Sasai, M. et al. (2010) Mol Immunol 47, 1283-91.
  23. Meylan, E. et al. (2004) Nat Immunol 5, 503-7.
  24. Jiang, Z. et al. (2004) Proc Natl Acad Sci USA 101, 3533-8.

Application References

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