| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
ODN-2395 specifically targets Toll-like receptor 9 (TLR9) within endosomes of plasmacytoid dendritic cells and B cells. As a Class C agonist, it contains both a phosphorothioate backbone and a palindromic sequence. It strongly induces the production of both pro-inflammatory cytokines (e.g., IL-6, TNF-alpha) and type I interferons (e.g., IFN-alpha), thereby stimulating both innate and adaptive immune responses.
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| ln Vitro |
In vitro, ODN-2395 activates TLR9 signaling pathways. Treatment of human peripheral blood mononuclear cells (PBMCs) with ODN-2395 leads to the maturation of plasmacytoid dendritic cells (pDCs) and B cells. It potently induces the secretion of a broad range of cytokines and chemokines, including IFN-alpha, IP-10, IL-6, and TNF-alpha. It also upregulates the expression of costimulatory molecules (CD80, CD86) on antigen-presenting cells.
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| ln Vivo |
In vivo, ODN-2395 is used as a vaccine adjuvant. When co-administered with an antigen (e.g., a viral protein or a tumor antigen), it significantly enhances the magnitude and quality of the antigen-specific immune response. It promotes a Th1-biased immune response, characterized by high levels of IgG2a antibodies and CD8+ T cell responses. Its efficacy as an adjuvant has been demonstrated in preclinical models of infectious disease and cancer.
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| Enzyme Assay |
For non-cell-based TLR9 binding assays, a standard protocol involves using biotinylated ODN-2395 and streptavidin-coated plates. Recombinant human or mouse TLR9 ectodomain is added to the plate. Binding is detected using an anti-TLR9 antibody. Competitive binding assays using unlabeled ODN-2395 or a non-specific ODN can be performed to assess selectivity. Alternatively, a surface plasmon resonance (SPR) assay can be used to determine the KD for the ODN-2395/TLR9 interaction.
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| Cell Assay |
For in vitro cell assays, mouse splenocytes or human PBMCs are isolated and seeded in 96-well plates. Cells are treated with ODN-2395 at concentrations ranging from 0.1-10 microM for 24-48 hours. Supernatants are collected and analyzed for cytokine secretion using a multiplex ELISA assay. Alternatively, the activation of NF-kappaB or IRF pathways can be measured using reporter cell lines (e.g., HEK-Blue hTLR9 cells) that secrete an alkaline phosphatase reporter upon TLR9 activation.
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| Animal Protocol |
For in vivo animal models, ODN-2395 is typically used as an adjuvant. A standard protocol involves immunizing mice (e.g., C57BL/6) subcutaneously or intramuscularly with a model antigen (e.g., OVA, 10 microg) mixed with ODN-2395 (e.g., 20-50 microg per mouse) on day 0 and day 14. A control group receives antigen alone. Blood is collected on day 28 to measure antigen-specific antibody titers (IgG, IgG1, IgG2a) by ELISA. Spleens are harvested for T-cell ELISpot assays to measure IFN-gamma secreting T cells.
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| ADME/Pharmacokinetics |
ODN-2395 is a 22-mer oligonucleotide with a molecular weight of approximately 7035 Da. Its phosphorothioate backbone confers resistance to nuclease degradation, giving it a relatively long plasma half-life compared to native DNA (estimated several hours in vivo). When used as an adjuvant, it is typically administered at the site of injection, where it is taken up by local immune cells. Systemic absorption leads to predictable immune activation, and it is eventually cleared through cellular metabolism.
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| Toxicity/Toxicokinetics |
As an immunostimulatory oligonucleotide, the toxicity profile of ODN-2395 is related to its mechanism of action. Overactivation of TLR9 can lead to a "cytokine storm," characterized by excessive release of inflammatory cytokines leading to fever, hypotension, and tissue damage. In preclinical studies, systemic administration may cause transient flu-like symptoms and splenomegaly (enlarged spleen). Chronic administration could theoretically lead to autoimmunity. It is a research tool and not a clinically approved drug.
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| References | |
| Additional Infomation |
ODN-2395 is a Class C CpG ODN, representing one of the three main classes (A, B, C) of TLR9 agonists distinguished by their structure and cytokine profile. Class A ODNs are strong inducers of IFN-alpha but weak inducers of B cell activation; Class B are vice versa; Class C ODNs like ODN-2395 induce a balanced response, making them particularly attractive as vaccine adjuvants. Its palindromic sequence allows it to form multimeric structures, which may be important for its high activity. It is not approved for human use but is widely used in veterinary vaccines and research.
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| Molecular Formula |
NA??MOLECULARWEIGHT
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|---|---|
| Molecular Weight |
0
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| CAS # |
1254617-22-5
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| Related CAS # |
Biotin-labeled ODN 2395 sodium;FITC-labeled ODN 2395 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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