Technical datasheet
Description
Bacterial DNA is known to induce acute inflammatory responses. Bacterial DNA acts as a pathogen-associated molecular pattern by virtue of a 20-fold greater frequency of unmethylated CG dinucleotides found in microbial DNA versus vertebrate DNA. Cellular activation by deoxy-cytidylate-phosphate-deoxy-guanylate (CpG)-DNA occurs via the Toll/IL-1R signal pathway. TLR9 appears to be a major component of the CpG-DNA receptor, acting by direct binding to CpG-DNA.
The synthetic oligodeoxynucleotides (ODN) contain the proper CpG-DNA motif mimicing the immunostimulatory effects of bacterial DNA. The human optimal CpG motif GTCGTT differs from the optimal mouse CpG motif GACGTT. Non CpG-DNA has been shown to compete with CpG-DNA in in vitro stimulation experiments.
This mouse CpG is a 20-mer ODN has the following sequence: 5'-tccatgacgttcctgatgct-3'. Regular letters represent phosphorothiorate linkage.
Formulation
Approximately 200 nmol lyophilized, purified 20 mer CpG oligodeoxynucleotide. The exact amount is
indicated on the label. Reconstitute the vial by injection of distilled or deionized water, volume depending
on the concentration to be used.
Application
CpG-DNA can be used in biological assays in vitro to activate murine cells.
Use
For stimulation in vitro 0.01 to 10 µM can be used. It is recommended that users test the reagent and determine their own optimal concentrations.
Storage and stability
Lyophilized product should be stored at 4°C. Store stock solution in aliquots at -70°C. Repeated freeze and thaw cycles will cause loss of activity. Under recommended storage conditions, product is stable for one year.
References
1. Bauer, S et al; Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Natl Acad Sci USA 2003, 98: 9237 2. Hemmi, H et al; A toll-like receptor recognizes bacterial DNA. Nature 2000, 408: 740
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Also available
References
1. Bauer, S et al; Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. Proc Natl Acad Sci USA 2003, 98: 9237
2. Hemmi, H et al; A toll-like receptor recognizes bacterial DNA. Nature 2000, 408: 740