ODN 2006 (ODN 7909)

Class B CpG oligonucleotide; a human TLR9 ligand

Catalog # tlrl-2006, tlrl-2006-1, tlrl-2006-5

For research use only

Version # 16E05-MM

3. Krug A. et al., 2001. Identification of CpG oligonucleotide sequences with high induction of IFN-alpha/beta in plasmacytoid dendritic cells. Eur J Immunol, 31:2154-1. Krieg, A. et al., 1995. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 374:546-9. 2. Bauer, S. et al., 2001. Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition. PNAS, 98:9237-42.

63. 4. Marshall J. et al., 2005. Superior activity of the type C class of ISS in vitro and in vivo across multiple species. DNA Cell Biol. 24(2):63-72.

METHODS

Preparation of stock solution (500 µM)

TLR9 activation can be achieved with 1-5 µM ODN 2006. • Resuspend ODN 2006 with endotoxin-free water (provided).

- Add 52 µl to 200 µg of ODN 2006 - Add 260 µl to 1 mg of ODN 2006

• Vortex until completely dissolved. Prepare aliquots and store at -20°C. • Prepare serial dilutions using endotoxin-free water.

^Note: The working concentration may vary depending on the levels of TLR9 gene expression and the species from which the gene was obtained.

TLR9 stimulation using ODN 2006

ODN 2006 can be used to stimulate TLR9 in HEK-Blue™ TLR9 cells. HEK-Blue™ TLR9 cells stably overexpress the TLR9 gene and an NF- B-inducible secreted embryonic alkaline phosphatase (SEAP) reporter genek. For more information, visit: www.invivogen.com

Below is a protocol to study TLR9 stimulation using HEK-Blue™ TLR9 cells in a 96-well plate.

- Dispense 20 l of stimulatory or control ODN per well of a 96-well plate. m

- Prepare cell suspension of HEK-Blue™ TLR9 cells according to the data sheet.

- Add HEK-Blue™ TLR9 cells (4-8 x104) to each ODN-containing well. - Incubate for 6-24 h at 37°C, 5% CO2.

- Determine TLR9 stimulation by assessing cytokine expression using ELISA, or SEAP expression using QUANTI-Blue™, a SEAP detection medium.

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