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| 1mg |
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| 2mg |
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| Targets |
ODN-2216 targets human Toll-like receptor 9 (TLR9). As a Class A (also known as type D) CpG ODN, it has a phosphodiester (PO) central palindromic CpG motif and a phosphorothioate (PS)-modified 3' poly-G tail. It strongly activates TLR9 to induce type I interferons (IFN-alpha, IFN-beta).
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| ln Vitro |
ODN 2216 (6 μg/mL, 9 days) stimulates naive CD4 T cell Th1 development [1]. Thymic DC cell apoptosis is greatly reduced by ODN 2216 (2 μM, 15 hours) [2].
In vitro, ODN-2216 (6 ug/mL, 9 days) stimulates naive CD4+ T cell Th1 development via an IFN-alpha/IL-12-dependent mechanism. It also greatly reduces apoptosis in thymic dendritic cells (2 uM, 15 hours, apoptosis rate 23.1% vs. control). It activates pDCs and cDCs to produce IL-12 and indirectly stimulates IFN-gamma release in PBMCs, enhancing NK cell and TCR-triggered CD4+ T cell responses. |
| ln Vivo |
In a mouse vaccination model, ODN 2216 stimulates T cell-mediated anti-tumor immune responses[3].
ODN-2216 induces T-cell mediated antitumor immune responses in murine vaccination models. It has also been used to study the mtDNA/TLR9/MicroRNA-223 negative feedback loop, which regulates neutrophil activation and protects against acetaminophen-induced hepatotoxicity in experimental animal models. |
| Enzyme Assay |
Not applicable. ODN-2216 is a TLR9 agonist oligonucleotide; its activity is measured in cell-based activation assays assessing TLR9-mediated signaling, including NF-kappaB activation, IRF activation, and downstream cytokine production.
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| Cell Assay |
Apoptosis Analysis[2]
Cell Types: Thymic DC, pDC Tested Concentrations: 2 μM Incubation Duration: 15 h Experimental Results: Dramatically diminished apoptosis of thymic DC (23.1 ± 0.9%), and naturally induced apoptosis of thymic pDC was not diminished (72.8 ± 2.9 %). Human PBMCs or purified immune cell subsets (pDCs, cDCs, T cells) are seeded. ODN-2216 is added at concentrations of 1-10 ug/mL for 6-48 h. Cytokines (IFN-alpha, IFN-beta, IL-12, IFN-gamma) are quantified by ELISA. pDC and cDC activation is assessed by flow cytometry (e.g., CD86, HLA-DR). T cell differentiation is analyzed by flow cytometry for Th1 markers (e.g., CXCR3, T-bet). |
| Animal Protocol |
In mouse models, ODN-2216 is used as an immunoadjuvant. It is typically administered via intraperitoneal or subcutaneous injection at 10-100 ug per mouse in combination with antigens to elicit T-cell mediated antitumor immune responses. For hepatotoxicity studies, it is administered before acetaminophen challenge to study the TLR9-mtDNA pathway.
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| ADME/Pharmacokinetics |
TK data are limited. ODN-2216 is soluble in H2O. It is formulated in endotoxin-free water or PBS for injection. Storage: powder at -20degC for 3 years; in solvent at -80degC for 1 year. Molecular weight: approx. 6432. Sequence: DNA, d(G-sp-G-sp-G-G-G-A-C-G-A-T-C-G-T-C-G-sp-G-sp-G-sp-G-sp-G-sp-G).
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| Toxicity/Toxicokinetics |
Standard oligonucleotide safety precautions apply. ODN-2216 is for research use only, not for human therapy. Avoid inhalation and contact. Use PPE in a well-ventilated area. The compound may induce strong immunostimulatory effects.
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| References | |
| Additional Infomation |
This is a research-grade human-specific TLR9 agonist. It is used to study antiviral immunity, cancer immunotherapy, and autoimmune diseases. ODN-2216 is also used as a positive control for type I interferon induction in human TLR9 research.
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| CAS # |
332437-00-0
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| Related CAS # |
FITC-labeled ODN 2216 sodium;Biotin-labeled ODN 2216 sodium
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| Appearance |
White to off-white solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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