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 (1-3). TLRs recognize conserved motifs found in various pathogens and mediate defense responses. Triggering of the TLR pathway leads to the activation of NF-kB and subsequent regulation of immune and inflammatory genes. The TLRs and members of the IL-1 receptor family share a conserved stretch of approximately 200aas known as the TIR domain. Upon activation, TLRs associate with a number of cytoplasmic adaptor proteins containing TIR domains including MyD88 (myeloid differentiation factor), MAL/TIRAP (MyD88-adaptor-like/TIR-associated protein),
TRIF (Toll-receptor-associated activator of interferon) and TRAM (Toll-receptor-associated molecule). This association leads to the recruitment and activation of IRAK1 and IRAK4,
which form a complex with TRAF6 to activate TAK1 and IKK. Activation of IKK leads to the degradation of IkB that normally maintains NF-kB inactivity by sequestering it in the cytoplasm.
TLR7, 8 and 9 form a group of structurally related TLR family members that are are localized to intracellular endosomes (4-6). TLR7 shows highest expression in lung, placenta, and spleen (4). TLR7 mediates responses to a class of synthetic compounds, including imidazoquinolines,
guanosine-based drugs that induce anti-viral responses (7). TLR7 responds to ssRNA viruses to activate NF-kB and trigger IFN production (8-10).
Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.
Recommended Dilution:
Western Blot: 1:1000 Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4 degrees C with gentle shaking, overnight.
Immunoprecipitation: 1:100
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.