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ODN 1720 (TLRgrade™) (synthetic) (Control)

Negative control for ODN 1668
 
ALX-746-200-T100 100 tests 178.00 USD
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GpC oligodeoxynucleotide with phosphorothioate backbone.

Product Specification

Sequence:5’-tccatgagcttcctgatgct-3’ (lower case letters indicate phosphorothioate linkage).
 
MW:6383 (ammonium salt)
 
Source:Synthetic.
 
Quantity:15nmol (~96µg). 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).
 
Formulation:Lyophilized. Sterile.
 
Purity Detail:Activity and endotoxin tested - TLRgrade™.
 
Endotoxin Content:<0.002 EU/µg (LAL test; BioWhittaker)
 
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.
 
Shipping:Ambient
 
Long Term Storage:+4°C
 
Use/Stability:Aqueous stock solution is stable for 1 day when stored at +4°C.
 
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.
 
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-746-001.
 
746-001 1
Figure 1: TLR9-dependent activation of mouse macrophages by ODN 1668 (TLRgrade™) (synthetic) (Prod. No. ALX-746-001). Method: Culture of 5x105 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).
746-001 2
Figure 2: TLR9-dependent mitogenic activity of ODN 1668 (TLRgrade™) (synthetic) (Prod. No. ALX-746-001) in mouse splenocytes. Method: Culture of 5x105 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 hours3H-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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