| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
TLR9 (Toll-like receptor 9). ODN 1018 signals through TLR9 to induce immunoglobulin production by B cells and interferon (IFN-alpha, IFN-beta), interleukin-12 (IL-12), and tumor necrosis factor-alpha (TNF-alpha) production by plasmacytoid dendritic cells (pDCs). As a Class B CpG ODN, it strongly activates B cells, promoting proliferation, antibody production (including IgG and IgM), and pro-inflammatory cytokine release (e.g., IL-6, TNF-alpha). Unlike Class A CpG ODNs, Class B ODNs weakly stimulate IFN-alpha secretion from pDCs but more potently activate B-cell responses. The FITC-labeled version retains TLR9 binding specificity and immune stimulatory activity.
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| ln Vitro |
In vitro, ODN 1018 induces strong B cell activation, proliferation, and immunoglobulin secretion in primary human B cell cultures. It promotes the production of pro-inflammatory cytokines including IL-6, TNF-alpha, and IL-12 from TLR9-expressing immune cells. In pDC cultures, ODN 1018 induces moderate levels of IFN-alpha, IFN-beta, IL-12, and TNF-alpha, contributing to Th1-biased immune responses. The compound also upregulates co-stimulatory molecules (CD80, CD86, MHC class II) on antigen-presenting cells, enhancing antigen presentation capability. The FITC-labeled version retains full cytokine-inducing activity, enabling functional studies to be performed simultaneously with uptake and localization assessments.
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| ln Vivo |
In animal models, ODN 1018 acts as a potent vaccine adjuvant, enhancing antigen-specific antibody responses when co-administered with vaccine antigens. In non-human primate studies, ODN 1018 promotes antigen presentation and co-stimulatory molecular expression on dendritic cells, leading to enhanced T-cell activation. In murine tumor models, ODN 1018 combined with immunotherapies such as rituximab shows anti-tumor activity through enhanced immune cell activation. In infectious disease models, ODN 1018 administered with hepatitis B surface antigen induces robust HBsAg-specific antibody titers and Th1-biased cellular immunity. In vivo administration leads to increased serum cytokine levels, which can be quantified by ELISA within 2-6 hours post-injection. FITC-labeled ODN 1018 can be used for ex vivo tissue distribution analysis in lymphoid organs after in vivo dosing.
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| Enzyme Assay |
For non-cellular TLR9 binding evaluation, an ELISA-based binding assay can be performed. Recombinant human TLR9 protein (50 uL at 2 ug/mL) is coated onto a 96-well plate in carbonate-bicarbonate buffer (pH 9.6) overnight at 4degC. After washing with PBST (PBS with 0.05% Tween 20) and blocking with 3% BSA for 1 hour at 37degC, serially diluted FITC-labeled ODN 1018 (0-2000 nM) is added and incubated for 2 hours at room temperature. Bound ODN is detected using an HRP-conjugated anti-FITC antibody followed by TMB substrate, measuring absorbance at 450 nm. Competitive binding assays can be performed by pre-incubating TLR9-coated plates with unlabeled ODN 1018 before adding labeled ODN. Alternatively, SPR can be used for real-time binding affinity measurement.
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| Cell Assay |
For cellular uptake and functional assessment, human PBMCs, purified B cells, or HEK293 cells stably expressing TLR9 are seeded in 96-well plates (2 × 10^5 cells/well) in RPMI-1640 containing 10% FBS. Cells are treated with FITC-labeled ODN 1018 at concentrations ranging from 0.1-20 uM for 2-48 hours. Cellular uptake and trafficking are assessed by confocal laser-scanning microscopy (excitation 495 nm, emission 520 nm) to visualize ODN accumulation in endosomal compartments where TLR9 is localized, or quantified by flow cytometry (FL1 channel). Functional activation is measured by collecting culture supernatants and quantifying cytokine secretion (IL-6, TNF-alpha, IFN-alpha, IL-12) via sandwich ELISA. B cell proliferation can be assessed by 3H-thymidine incorporation or CFSE dilution assay. Co-incubation with the TLR9 antagonist ODN 2088 serves as a specificity control.
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| Animal Protocol |
For in vivo distribution studies, animals (mice or rats) receive a single intravenous (tail vein), intraperitoneal, or subcutaneous injection of FITC-labeled ODN 1018 at doses ranging from 10-100 mg/kg. At predetermined time points (0.5, 1, 2, 6, 12, 24, 48, 72 hours post-injection), blood is collected and lymphoid tissues (spleen, lymph nodes, bone marrow), liver, kidneys, and lungs are harvested. Tissues are homogenized in lysis buffer, and FITC fluorescence (excitation 485 nm, emission 528 nm) is measured using a fluorescence plate reader, with values normalized to tissue weight. For histology, frozen tissue sections (8-10 um) are counterstained with DAPI and examined by fluorescence microscopy to determine cellular and subcellular distribution patterns. For vaccine adjuvant studies, animals are co-injected with FITC-labeled ODN 1018 and antigen; the draining lymph nodes are harvested at 24-48 hours post-injection for analysis of antigen uptake by dendritic cells and germinal center B cell activation.
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| ADME/Pharmacokinetics |
Class B CpG ODNs such as ODN 1018 exhibit improved nuclease resistance due to their full phosphorothioate backbone, resulting in a plasma half-life of approximately 1-2 hours in mice following intravenous administration, significantly longer than Class A CpG ODNs with phosphodiester backbones. The compound distributes extensively to lymphoid tissues including spleen, lymph nodes, and Peyer's patches, with peak tissue concentrations achieved within 1-2 hours post-injection. Liver and kidney uptake also occurs due to reticuloendothelial system clearance, but to a lesser extent than Class A ODNs. The full phosphorothioate backbone also promotes binding to plasma proteins, reducing renal clearance. Urinary excretion of metabolites is the primary elimination pathway. The FITC label does not significantly alter the pharmacokinetic properties of the parent ODN.
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| Toxicity/Toxicokinetics |
In preclinical toxicity studies, ODN 1018 is generally well-tolerated at immunomodulatory doses. At high doses or with frequent administration, cytokine-mediated adverse effects may occur including transient fever, malaise, lymph node enlargement, and mild elevation of liver transaminases, consistent with the class effects of TLR9 agonists. In chronic repeat-dose toxicity studies in rodents and non-human primates, no target organ toxicity, genotoxicity, or carcinogenicity has been reported at therapeutic dose levels. Lymphoid hyperplasia in spleen and lymph nodes is expected as a pharmacodynamic effect rather than an adverse toxicity. The FITC label does not introduce additional toxicological concerns beyond those of the parent ODN. Standard dose-escalation and tolerability assessments should be performed when transitioning to new animal models.
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| References | |
| Additional Infomation |
FITC-labeled ODN 1018 is a research tool for studying CpG ODN cellular uptake, endosomal trafficking to TLR9-containing compartments, and TLR9-mediated B cell activation. The unlabeled ODN 1018 has been investigated in clinical research as a vaccine adjuvant, notably in HEPLISAV™ (hepatitis B vaccine) where it serves as an immunostimulatory adjuvant, and in Phase II studies for relapsed or refractory follicular lymphoma in combination with rituximab (NCT00490568). ODN 1018 has not received regulatory approval as a standalone therapeutic agent; its clinical use is as a component of adjuvanted vaccines. The FITC label enables real-time visualization and quantification of ODN distribution using fluorescence-based techniques such as confocal microscopy and flow cytometry.
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| Molecular Weight |
8171.00
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| Related CAS # |
ODN 1826;202668-42-6;Biotin-labeled ODN 1018 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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.1224 mL | 0.6119 mL | 1.2238 mL | |
| 5 mM | 0.0245 mL | 0.1224 mL | 0.2448 mL | |
| 10 mM | 0.0122 mL | 0.0612 mL | 0.1224 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
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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