Cell Signaling Technology

Product Pathways - NF-kB Signaling

Toll-like Receptor 9 (D2C9) Rabbit mAb #5845

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H Endogenous 130 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human
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 9 (D2C9) Rabbit mAb recognizes endogenous levels of total TLR9 protein. This antibody is predicted to react with known full-length isoforms of TLR9, but not with the cleaved TLR9 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with recombinant protein specific to the central ectodomain of human TLR9 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Toll-like Receptor 9 (D2C9) 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.

TLR9 is highly expressed in macrophages, dendritic cells, and B lymphocytes, and in humans has five isoforms generated by alternative splicing (4,5). TLR9 binds to unmethylated CpG motifs present on bacterial DNA and stimulates NF-κB via the MyD88 adaptor protein (6-8). In contrast to most TLR family members that are localized to the plasma membrane, TLR9 is an intracellular receptor localized to the ER in resting cells (9). Upon binding to CpG DNA, TLR9 is proteolytically processed and translocates to endo-lysosomal compartments where it binds MyD88, initiating downstream signaling (10-12).

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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. Du, X. et al. (2000) Eur Cytokine Netw 11, 362-71.
  16. Chuang, T.H. and Ulevitch, R.J. (2000) Eur Cytokine Netw 11, 372-8.
  17. Hemmi, H. et al. (2000) Nature 408, 740-5.
  18. Bauer, S. et al. (2001) Proc Natl Acad Sci U S A 98, 9237-42.
  19. Takeshita, F. et al. (2001) J Immunol 167, 3555-8.
  20. Latz, E. et al. (2004) Nat Immunol 5, 190-8.
  21. Park, B. et al. (2008) Nat Immunol 9, 1407-14.
  22. Ewald, S.E. et al. (2008) Nature 456, 658-62.
  23. Sepulveda, F.E. et al. (2009) Immunity 31, 737-48.

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