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  7. CpG ODN (1826), TLR9 ligand (Class B)

CpG ODN (1826), TLR9 ligand (Class B)

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Fig-1: Induction of mouse TLR9 activity by CpG ODN 1826.

CpG ODN (1826), TLR9 ligand (Class B)
CpG ODN (1826), TLR9 ligand (Class B)
CpG ODN (1826), TLR9 ligand (Class B)

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Fig-1: Induction of mouse TLR9 activity by CpG ODN 1826.
Fig-2: Abeomics' CpG ODNs did not show any TLR2 agonist activity. TLR2/NF-kB Leeporter™ HEK293 cells (14-127ACL) were treated with various CpG ODNs at 100 ug/ml as well as a positive TLR2 agonist, Pam3CSK4, at 10 ng/ml for 16 h, and luciferase activity was then analyzed.
Fig-3: Abeomics' CpG ODNs did not show any TLR4 agonist activity. TLR4/IL-8 Leeporter™ HeLa cells (14-124ACL) were treated with various CpG ODNs at 100 ug/ml as well as a positive TLR4 agonist, LPS, at 10 ng/ml for 16 h, and luciferase activity was then analyzed.

Product code: 15-1018

Application : Functional Assay
Reactivity : Rat, Mouse

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Amount : 0.1 mg
Content : 1 mg/ml in entotoxin-free water
Storage condition : Upon receipt, store at -20°C (Stable for at least 6 months). Avoid frequent freeze/thaw cycles.
Sequence: 5'-tccatgaCGttcctgaCGtt-3' (Class B ODN 1826: Bases are phosphorothioate.)
 
Synthetic oligodeoxynucleotides (ODN) containing unmethylated deoxycytosine-deoxyguanosine (CpG) motif are equivalent to bacterial DNA in the immunostimulatory activity, which can induce innate immunity via Toll-like receptor 9 (TLR9) in mammals. There are three major classes of CpG ODNs, and each ODN class exhibits different stimulatory effects on immune cell activation. Class A ODNs are potent inducers of IFN-alpha that leads to the plasmacytoid dendritc cell (pDC) maturation. Class B ODNs are relatively weak inducers of type I IFNs but strong stimulators of human B cells and monocyte maturation. Class C ODNs combine elements of both Classes A and B ODNs, which can induce IFN-alpha in pDC and activation of B cells.

B. CpG ODN 1826-mediated mouse TLR9 activation in NF-kB Leeporter™ – RAW 264.7 cells (Figure 2). 
 
1. Harvest NF-kB Leeporter™ – RAW 264.7 cells and seed cells into a white solid-bottom 96-well microplate in 100 ul of growth medium at 5 x 104 cells/well.
 
2. Incubate cells at 37oC in a CO2 incubator for overnight.
 
3. The next day, stimulate cells with various amounts of CpG ODN 1826. 
 
4. Incubate at 37oC in a CO2 incubator for 6-16 hours.
 
5. Add 30-50 ul of luciferase assay reagent per well. 
 
6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
 

 

For Research Use Only. Not for use in diagnostic/therapeutics procedures.

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CpG ODN (1826), TLR9 ligand (Class B)

Product code: 15-1018
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