Toll-like receptors (TLR) are highly conserved throughout evolution and have been implicated in the innate defence to many pathogens. In Drosophila toll is required for the anti-fungal response, while the related 18-wheeler is involved in antibacterial defences. In mammals, TLR identified as type I transmembrane signalling receptors with pattern recognition capabilities, have been implicated in the innate host defence to pathogens. As investigated so far all functional characterized TLR signal via the TLR/IL-1 receptor (IL-1R) pathway where recruitment of MyD88 seems to be essential. In contrast to cell-wall components, bacterial DNA is probably invisible for immune cells until DNA is liberated during processes taking place in the endosomal/lysosomal compartment where intracellular TLR9 (CD289) recruits MyD88 to initiate signal transduction. Unmethylated CpG-dinucleotide-containing sequences are found much more frequently in bacterial genomes than in vertebrates genomes, whereas the frequency of CpG dinucleotides are suppressed and usually methylated. The regions adjacent to the CpG dinucleotides also affect the immunostimulatory activity. The optimal sequence differs significantly between mammalian species. Methylated CpG dinucleotides lack immunostimulatory activities. Cellular activation in response to bacterial DNA and synthetic dinucleotides containing unmethylated CpG-dinucleotides is mediated by TLR9.
Applications:
Can be used for Flow Cytometry, Western Blot, ELISA (detection), and Immunohistochemistry (frozen). Stains RAW macrophages and TLR9 (CD289) transfected HEK293 cells. Other applications not tested.
Recommended Dilution:
Immunohistology: 1:50
Flow Cytometry: 1:50
Western Blot: 1:50
Optimal dilutions to be determined by the researcher.
Storage and Stability:
For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. FITC conjugates are sensitive to light.