CpG ODN (1585), TLR9 ligand (Class A)

Product code: 15-1020

Application : Functional Assay
Reactivity : Mouse, Human

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  •   1 mg

  •  0.1 mg

  • $409.00 

  • $130.00 

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Shipping Info:

Order now and get it on Wednesday June 26, 2024

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Amount : 0.1 mg
Content : 1 mg/ml in endotoxin-free water
Storage condition : Upon receipt, store at -20°C (Stable for at least 6 months). Avoid frequent freeze/thaw cycles.
Sequence: 5'-ggggtcaaCGttgagggggg-3' (Class A ODN 1585: 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.

A. CpG ODN 1585-mediated human TLR9 activation in TLR9/NF-kB Leeporter™ – HEK293 cells (Figure 2). 
 
1. Harvest TLR9/NF-kB Leeporter™ – HEK293 cells and seed cells into a white solid-bottom 96-well microplate in 100 µl 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 1585. 
 
4. Incubate at 37oC in a CO2 incubator for 6-16 hours.
 
5. Add 30-50 µl of luciferase assay reagent per well. 
 
6. Incubate at room temperature for 1-5 minutes and measure luminescence using a microplate luminometer.
 
 
B. CpG ODN 1585-mediated mouse TLR9 activation in NF-kB Leeporter™ – RAW 264.7 cells (Figure 3). 
 
1. Harvest NF-kB Leeporter™ – RAW 264.7 cells and seed cells into a white solid-bottom 96-well microplate in 100 µl 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 1585. 
 
4. Incubate at 37oC in a CO2 incubator for 6-16 hours.
 
5. Add 30-50 µl 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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