Cell Signaling Technology

Product Pathways - NF-kB Signaling

Toll-like Receptor Antibody Sampler Kit #9971

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Toll-like Receptor 1 Antibody #2209 40 µl W IP H (Mk) 86 Rabbit
Toll-like Receptor 7 Antibody #2633 40 µl W H M R (Mk) (Dg) 140 Rabbit
Toll-like Receptor 2 Antibody #2229 40 µl W H Mk 90-105 Rabbit
Toll-like Receptor 9 Antibody #2254 40 µl W IP H 130 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dg=Dog
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

All antibodies contained in this kit detect endogenous levels of their respective total TLR protein. Cross reactivity was not detected with other family members at physiological conditions.

Western Blotting

Western Blotting

Western blot analysis of extracts from SR cells using Toll-like Receptor 1 Antibody #2209.

Western Blotting

Western Blotting

Western blot analysis of extracts from THP-1 cell lines, stimulated with 200 µM TPA overnight, using Toll-like Receptor 2 Antibody #2229.

Western Blotting

Western Blotting

Western blot analysis of extracts from Raji and Ramos cells using Toll-like Receptor 9 Antibody #2254.


Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, mock transfected or transfected with human TLR7, and from Ramos and A20 cell lines using Toll-like Receptor 7 Antibody #2633.

Description

The Toll-Like Receptor Antibody Sampler Kit is an economical way to examine the total protein levels of a number of toll-like receptors. This kit includes enough primary and secondary antibodies to perform four Western blot experiments with each antibody.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide and the resulting antibody is purified by peptide affinity chromatography. The Toll-like Receptor 1 Antibody #2209 is specific for the amino terminus of human TLR1. The Toll-like Receptor 2 Antibody #2229 is specific for the region surrounding leucine 179 of human TLR2. The Toll-like Receptor 7 Antibody #2633 is specific for the carboxyl terminus of human TLR7. The Toll-like Receptor 9 Antibody #2254 is specific for the region surrounding glycine 442 of human TLR9.

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.

  1. Akira, S. (2003) J Biol Chem 278, 38105-8.
  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.

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