Cell Signaling Technology

Product Pathways - NF-kB Signaling

TLR8 (D3Z6J) Rabbit mAb #11886

Applications Reactivity Sensitivity MW (kDa) Isotype
W H (Mk) Endogenous 150 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

TLR8 (D3Z6J) Rabbit mAb recognizes endogenous levels of total TLR8 protein. This antibody cross-reacts with a 30 kDa protein and a 37 kDa protein of unknown origin.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro141 of human TLR8 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cells differentiated with TPA #4174 (80 nM, 16 hr), untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (20 ng/mL, 8 hr; +), using TLR8 (D3Z6J) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing HA-tagged full-length human TLR8 (hTLR8-HA; +), using TLR8 (D3Z6J) 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.

TLR8 is an intracellular TLR localized to the endoplasmic reticulum, endosomes, lysosomes, and endolysosomes (4). It is activated by single-stranded viral RNA, as well as synthetic imidazoquinoline compounds including R-848 (5). TLR8 expression is highest in the lung and in myeloid cells (6,7). In addition, expression is upregulated by IFN-γ in monocyte-like leukemic THP-1 cells that have been differentiated with TPA (7).

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  2. Beutler, B. (2004) Nature 430, 257-63.
  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. Heil, F. et al. (2003) Eur J Immunol 33, 2987-97.
  16. Jurk, M. et al. (2002) Nat Immunol 3, 499.
  17. Chuang, T.H. and Ulevitch, R.J. (2000) Eur Cytokine Netw 11, 372-8.
  18. Zarember, K.A. and Godowski, P.J. (2002) J Immunol 168, 554-61.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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