Anti-Mouse CD134 (Clone OX-86) – Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD134 (Clone OX-86) – Purified in vivo PLATINUM™ Functional Grade

Product No.: C856

[product_table name="All Top" skus="C392"]

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Clone
OX-86
Target
CD134
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
OX-40, TNFRSF4, ACT35
Isotype
Rat IgG1
Applications
Act
,
FC
,
IHC
,
in vivo
,
WB

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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Additional Applications Reported In Literature ?
FC
IHC
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Clone OX-86 reacts with murine CD134 (OX-40, TNFRSF4).
Background
CD134 functions as an important immune checkpoint, and its depletion in murine mouse models demonstrate that lack of CD134 expression leads to reduced CD4+ and CD8+ T cells1. When CD134 is bound by its corresponding ligand (OX-40L), an optimal T cell response is generated and plays a significant role in determining the amount of memory T-cells remaining after the immune response1. CD134 has also been found to play an important role in carcinogenesis, as treatment with activating in vivo antibodies against CD134 enhanced tumor growth, suggesting that CD134 is an important tumor suppressor, and its absence disrupts the immune response to tumors2,3.
Antigen Distribution
CD134 is expressed on activated CD4 and CD8 T cells, activated regulatory T cells, B cells, NKT cells, NK cells, and neutrophils.
Research Area
Immunology

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Clone OX-86 is a monoclonal antibody that targets mouse OX40 (CD134), acting as an agonist to stimulate the OX40 pathway. Common in vivo applications of clone OX-86 in mice include:

  • Enhancing T Cell Responses: OX-86 stimulates the OX40 pathway, which is crucial for enhancing T cell clonal expansion and cytokine production. This is particularly beneficial in immunotherapy strategies aiming to boost immune responses against pathogens or tumors.

  • Modulating Immune Regulation: By activating the OX40 pathway, OX-86 can influence the balance between effector and regulatory T cells, potentially affecting immune tolerance and autoimmunity.

  • Cancer Research: The agonistic effects of OX-86 have been used to delay tumor growth in mice by enhancing the generation of antigen-specific effector T cells and preventing T cell tolerance.

  • Vaccine Efficacy Studies: OX-86 can be used to study the immune response to vaccines by augmenting the proliferation of antigen-reactive T cells, thereby improving vaccine efficacy.

Commonly Used Antibodies and Proteins with OX-86 in the Literature

OX-86 is a well-established monoclonal antibody against mouse CD134 (OX40/TNFRSF4), primarily used for studying T cell activation, immune checkpoint modulation, and the dynamics of OX40 signaling in various immunological contexts. In published studies, OX-86 is often used in combination with other antibodies and proteins to provide a comprehensive profile of T cell function, activation status, and immune regulation.

Key Antibodies Commonly Used with OX-86

Antibody/ProteinTargetPurposeExample Use with OX-86
Anti-IFN-γInterferon-gamma (IFN-γ)Measures Th1 cytokine production in T cellsCytokine detection assays (ELISA, intracellular staining)
Anti-IL-2Interleukin-2 (IL-2)Assesses T cell proliferation and activationCytokine detection assays
Anti-IL-4Interleukin-4 (IL-4)Detects Th2 cytokine responsesCytokine detection assays
Anti-IL-17AInterleukin-17A (IL-17A)Evaluates Th17 responsesCytokine detection assays
Anti-CD4CD4Identifies helper T cellsFlow cytometry, co-staining to identify CD4+ T cell subsets
Anti-CD25CD25/IL-2RαMarks activated and regulatory T cellsFlow cytometry, especially for Treg studies
CD252 (OX40L)OX40 ligandEngagement with OX40/CD134 to study OX40-OX40L signaling axisFunctional studies of OX40 signaling
Isotype controlsN/ANegative controls for antibody specificityUsed alongside OX-86 to ensure staining specificity

Commonly Used Proteins

  • OX40L-fusion proteins: These include engineered proteins such as MBL-OX40L and Fc-scOX40L, which directly bind OX40 (CD134) and are used in functional assays to verify OX40-binding specificity and activate OX40 signaling pathways.
  • FcγRIIb and FcγRIII proteins: These are relevant in understanding the effector function and in vivo activity of OX-86, as the rat IgG1 isotype of OX-86 interacts with mouse FcγRIIb and FcγRIII receptors, influencing its agonistic or regulatory effects.

Application Context

  • Flow Cytometry: OX-86 is often combined with antibodies like anti-CD4 and anti-CD25 to phenotype activated T cells, regulatory T cells, and memory T cells.
  • Cytokine Detection (ELISA, ICS): Used with antibodies against IFN-γ, IL-2, IL-4, and IL-17A to assess the effector functions of T cells following OX-86 stimulation or CD134 engagement.
  • Functional Assays: OX40L-fusion proteins are used to activate or block OX40 signaling, often compared with OX-86 in dose-dependent binding studies to confirm receptor specificity.
  • In Vivo Studies: OX-86 is used alongside isotype controls to evaluate therapeutic effects in mouse models of cancer and autoimmunity.

Conclusion

OX-86 is most frequently used in concert with antibodies targeting key cytokines (IFN-γ, IL-2, IL-4, IL-17A), T cell markers (CD4, CD25), and the OX40 ligand (CD252). Fusion proteins (e.g., OX40L-fusion constructs) and Fc receptor proteins are also used to probe the biology of the OX40 signaling axis in functional assays. This multi-parameter approach enables researchers to dissect complex T cell responses in the context of OX40 engagement.

Clone OX-86 is a monoclonal antibody targeting murine CD134 (OX-40, TNFRSF4) that has generated significant findings across immunology and cancer research. This antibody acts as an agonist, stimulating OX-40 signaling and has been extensively used to investigate T cell responses and anti-tumor immunity.

T Cell Costimulation and Cytokine Production

OX-86 demonstrates potent costimulatory effects on T cells when combined with TCR/CD3 stimulation. Studies show that OX-86 significantly increases IL-2 and IFN-γ production from both CD4+ and CD8+ T cells in a dose-dependent manner. This costimulatory activity enhances T cell proliferation and cytokine secretion, suggesting that OX-40 signaling amplifies the immune response beyond primary TCR activation alone.

Role in T Cell Memory Formation

When CD134 is bound by its corresponding ligand (OX-40L) or agonistic antibodies like OX-86, an optimal T cell response is generated that plays a significant role in determining the amount of memory T cells remaining after the immune response. In vivo treatment with OX-86 has been shown to strongly enhance the generation of antigen-specific effector T cells and prevent the induction of T cell tolerance. This finding has important implications for vaccine development and immunotherapy strategies.

Anti-Tumor Activity

Research demonstrates that OX-86 can delay tumor growth in vivo. However, the mechanism appears complex—treatment with activating in vivo antibodies against CD134 enhanced tumor growth in some contexts, suggesting that CD134 functions as an important tumor suppressor, and its absence disrupts the immune response to tumors. The anti-tumor effects likely depend on the specific tumor microenvironment and the balance of immune cell populations present.

NK Cell Enhancement

Beyond T cells, OX-86 engagement enhances natural killer (NK) cell function. Studies show that CD134 engagement on NK cells with an agonistic antibody increased antibody-dependent cellular cytotoxicity (ADCC) capacity and IFNγ secretion. This finding expands the therapeutic potential of OX-40 targeting beyond conventional T cell responses.

Isotype-Dependent Mechanisms

The biological activity of OX-86 varies significantly based on its isotype. The original rat IgG1 version interacts primarily with FcγRIIb and FcγRIII, giving it a low activatory-to-inhibitory FcγR binding ratio and the capacity for direct agonism through FcγRIIb-mediated crosslinking. In contrast, chimeric versions with mouse IgG2a constant regions interact strongly with all activatory FcγR and likely mediate effects through cellular depletion mechanisms rather than pure agonism.

Dosing regimens of clone OX-86 (anti-mouse OX40 mAb) vary by mouse model and experimental aim, commonly ranging from 10 µg to 250 µg per injection, with schedules from a single dose to repeated daily or weekly injections.

  • Syngeneic Tumor Models: For the CT26 colon carcinoma model, 100 µg of OX-86 is administered twice per week. This dose has been shown to activate immune responses and alter tumor growth dynamics.
  • Regulatory T Cell Studies: Naïve foxp3gfp reporter mice typically receive 0.25 mg (250 µg) intraperitoneally (i.p.) for 3 to 7 consecutive days to study impacts on Treg expansion and memory T cell generation.
  • Combined Immunotherapy: In combination regimens (e.g., with anti-CTLA4 and anti-CD137), doses as low as 10 µg or 30 µg per injection are used biweekly or at set intervals; 10 µg in triple-therapy showed efficacy with minimal toxicity, while higher frequencies (e.g., 5–6 injections of 30 µg) can cause treatment-related deaths in certain models like MC38.
  • Weight-Based Dosing: Some protocols use weight-adjusted dosing, such as 5 mg/kg, reflecting variation adapted for animal size and experimental goals.
  • General Guidance: Manufacturer and supplier protocols support dosing regimens between 10–250 µg/injection, emphasizing the need to tailor dose and frequency to the studied model and endpoint.

Toxicity and reaction risks increase at higher doses and more frequent administrations, including potential anaphylaxis with repetitive dosing.

Summary table:

Mouse Model / AimDose per InjectionFrequencyNotes
Syngeneic tumor (CT26)100 µg2× weeklyMonotherapy
Treg induction (foxp3gfp reporter mice)0.2–0.25 mg (200–250 µg)3–7 daily injectionsi.p. administration
Dual-tumor (MC38, A20) multi-therapy10 or 30 µg4 biweekly injectionsHigher doses/frequencies – toxicity
Weight-based5 mg/kgProtocol-specificFor combination therapies
General range across studies10–250 µgSingle–multiple weeklyMust be optimized by model

In conclusion, the OX-86 clone dosing regimen is highly context-dependent, with lower doses favored for combination therapies to minimize toxicity, and higher/frequent doses used for robust immune activation or mechanistic endpoints. Reviewing individual study protocols and monitoring for adverse reactions, especially at higher frequencies or doses, is recommended.

References & Citations

1. Redmond WL, Ruby CE, Weinberg AD. Crit Rev Immunol. 29(3):187-201. 2019
2. Morris A, Vetto JT, Ramstad T, et. al. Breast Cancer Res Treat. 67: 71–80. 2001.
3. Weinberg AD, Rivera MM, Prell R, et. al. J Immunol. 164: 2160–9. 2000.
4. al-Shamkhani A, Birkeland ML, Puklavec M, et. al. Eur J Immunol. Aug;26(8):1695-9. 1996.
5. Higgins LM, McDonald SA, Whittle N, et. al. J Immunol. Jan 1;162(1):486-93. 1999.
Act
Flow Cytometry
IHC
in vivo Protocol
General Western Blot Protocol

Certificate of Analysis

Formats Available

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.