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

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

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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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Download TDS / Manual MSDS

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 37^oC in a CO₂ incubator for overnight.

3. The next day, stimulate cells with various amounts of CpG ODN 1826.

4. Incubate at 37^oC 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.

Zhang K, Zakeri A, Alban T, Dong J, Ta HM, Zalavadia AH, Branicky A, Zhao H, Juric I, Husich H, Parthasarathy PB, Rupani A, Drazba JA, Chakraborty AA, Ching-Cheng Huang S, Chan T, Avril S, Wang LL.
VISTA promotes the metabolism and differentiation of myeloid-derived suppressor cells by STAT3 and polyamine-dependent mechanisms.
Cell Rep. 2024 Jan 3;43(1):113661. doi: 10.1016/j.celrep.2023.113661. Epub ahead of print.
PMID: 38175754.

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