For a larger pack size, please see Prod. No.
ALX-746-051.
CpG oligodeoxynucleotide (Type B) with phosphorothioate backbone. Specific ligand for mouse TLR9 (Toll-like receptor 9).
Product Details
Sequence: | 5’-tccatgacgttcctgatgct-3’ (lower case letters indicate phosphorothioate linkage). |
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MW: | 6383 (ammonium salt) |
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Source: | Synthetic. |
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Quantity: | 100µg (~15nmol). Sufficient for at least 100 cellular activation assays at 0.1-0.5µg/well when performed in a 200µl assay volume (i.e. 96 well microtiter plate). |
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Formulation: | Lyophilized. Sterile. |
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Purity Detail: | Activity and endotoxin tested - TLRGRADE®. |
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Endotoxin Content: | <0.002EU/µg (LAL test; BioWhittaker) |
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Reconstitution: | For a 100µM stock solution, dissolve the total vial content in 150µl endotoxin-free water (included) or PBS (to order separately). To obtain optimal dissolving we recommend the following procedure:
- Add 50% of the solvent and let dissolve for 10 min.
- Add remaining 50% of the solvent and mix thoroughly.
- Moderate warming may aid dissolving. |
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Shipping: | Ambient |
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Long Term Storage: | +4°C |
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Use/Stability: | Aqueous stock solution is stable for 1 day when stored at +4°C. |
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Handling: | Protect from light. For maximum product recovery after thawing, centrifuge the vial before opening the cap. After reconstitution, prepare aliquots and store at -20°C. |
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Technical Info/Product Notes: | Includes 1 vial of ddWater (endotoxin-free) (Prod. No. ALX-505-008). For inactive control compound, please see Prod. No. ALX-746-200. |
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Regulatory Status: | RUO - Research Use Only |
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Figure 1: TLR9-dependent activation of mouse macrophages by ODN 1668 (TLRGRADE®) (synthetic) (Prod. No.
ALX-746-001).
Method: Culture of 5x10
5 cells per 200µl well was performed in serum-free conditions after addition of ODNs at the indicated concentrations. Cell activation was measured by IL12-p40 induction. Only CpG ODN 1668 (Prod. No.
ALX-746-001), but not the corresponding GpC control ODN 1720 (Prod. No.
ALX-746-200) activates wild type mouse macrophages. Activation is absolutely dependent on the presence of TLR9 as addressed with cells derived from TLR9-deficient mice. Note the logarithmic scale for the x and y-axis and the high activity of the active ODNs: max. stimulation is already achieved with 0.010-0.100nmol per 200µl well (10-100nM, corresponding to ~0.5-5µg/ml).
Figure 2: TLR9-dependent mitogenic activity of ODN 1668 (TLRGRADE®) (synthetic) (Prod. No.
ALX-746-001) in mouse splenocytes.
Method: Culture of 5x10
5 cells per 200µl well was performed in serum-free conditions after addition of ODNs at the indicated concentrations. Cell proliferation was analyzed after 72 hours with a 16 hours
3H-thymidine pulse.Only CpG ODN 1668 (Prod. No.
ALX-746-001), but not the corresponding GpC control ODN 1720 (Prod. No.
ALX-746-200) induces proliferation of wild type mouse splenocytes. Induction of cell proliferation is absolutely dependent on the presence of TLR9 as addressed with cells derived from TLR9-deficient mice. Note the logarithmic scale for the x and y-axis and the high activity of the active ODNs: max. stimulation is already achieved with 0.010-0.100nmol per 200µl well (10-100nM, corresponding to ~0.5-5µg/ml).
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Product Literature References
Distinct effects of different adjuvants in the mouse model of allergic airway inflammation: H.S. Shin, et al.; Asian Pac. J. Allergy Immunol.
40, 111 (2022),
Abstract;
Paneth Cells Regulate Lymphangiogenesis under Control of Microbial Signals during Experimental Portal Hypertension: M. Hassan, et al.; Biomedicines
10, 1503 (2022),
Abstract;
Ly6C+ monocyte efferocytosis and cross-presentation of cell-associated antigens: S.R. Larson, et al.; Cell Death Differ.
23, 997 (2016),
Application(s): Efferocytosis assay, in vivo studies,
Abstract;
Full Text
Activation of murine macrophages via TLR2 and TLR4 is negatively regulated by a Lyn/PI3K module and promoted by SHIP1: S. Keck, et al.; J. Immunol.
184, 5809 (2010),
Abstract;
Full Text
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