Anti-Mouse CD96 – Purified in vivo PLATINUM™ Functional Grade

Anti-Mouse CD96 – Purified in vivo PLATINUM™ Functional Grade

Product No.: C780

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Clone
3.3
Target
CD96
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
Tactile (T cell-activated increased late expression)
Isotype
Rat IgG1 κ
Applications
B
,
FC
,
in vivo

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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.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM™ antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
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
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for this clone 3.3 antibody (anti-mouse CD96) for staining cells in flow cytometry is ≤ 1.0 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
Additional Applications Reported In Literature ?
B
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 3.3 recognizes an epitope on mouse CD96.
Background
CD96 is a single pass type I transmembrane glycoprotein in the immunoglobulin superfamily that is heavily N-glycosylated1. Murine (m) CD96 is present at the surface of most lymphocytes, including NK, CD4+ T, CD8+ T, NKT, and γδ T cells, but not B lymphocytes, neutrophils, macrophages, or dendritic cells2. mCD96 interacts with mCD155 and nectin-1 (CD111)1. A V-like domain mediates binding of mCD96 to mCD155 via interaction between amino acids of the FG loop of one binding partner with residues in the C’C’’-loop of the other. CD96 is a member of an interaction network that includes adhesion, activation, and inhibition activities.

CD96 contains three Ig-like domains that are separated from the transmembrane domain by a long proline, serine, and threonine rich stalk that undergoes extensive O-linked glycomodification1. The stalk may play a role in orientation or presentation of the Ig-like domains. mAb 3.3 binds to the first Ig domain and competes with CD155 for binding3.

Human CD96 has a mild boosting effect on 2B4- and NKp30-mediated killing, but a direct role in the activation of NK cell-mediated cytotoxicity in vitro has not been observed 1. In contrast, mCD96 suppresses NK cells in vivo2. Blocking studies show that mCD96 competes with CD226 for CD155 binding and limits NK cell function by direct inhibition2. Additionally, blocking mCD96 in vivo with mAb 3.3 protects against metastasis in three different tumor models. The antimetastatic effect of mAb 3.3 is independent of antibody-dependent cell-mediated cytotoxicity and activating Fc receptors3,4 and is enhanced by anti-PD-1 and anti-CTLA-4 mAbs4. Suppression of metastasis by mAb 3.3 is dependent on NK cells, CD226 (DNAM-1), and IFN-γ4. Additionally, mAb 3.3 loses its antimetastatic function in CD155- and IL-12p35-deficient mice3.
Antigen Distribution
CD96 (aka Tactile; T cell-activated increased late expression) is mainly expressed by cells of hematopoietic origin, in particular T cells and NK cells.
Ligand/Receptor
CD155, nectin 1
NCBI Gene Bank ID
Research Area
Immunology
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Inhibitory Molecules

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 3.3 is a monoclonal antibody targeting mouse CD96, and its most common in vivo applications in mice include functional studies of immune modulation, particularly focusing on natural killer (NK) cells and T cells.

Essential applications include:

  • Immune checkpoint studies: Clone 3.3 is commonly used to investigate the role of CD96 as an immune checkpoint receptor, especially regarding its inhibitory effects on NK cell activity. By blocking CD96 in vivo, researchers can assess changes in immune surveillance and anti-tumor responses.

  • Tumor immunology: The antibody is employed in models of cancer to determine whether CD96 blockade can enhance NK cell-mediated tumor rejection or modulate T cell responses within the tumor microenvironment. This is based on the role of CD96 in controlling cytotoxicity and cytokine production by immune cells.

  • Adoptive transfer and functional studies: It is used in combination with tumor challenge or adoptive cell transfer experiments to delineate how targeting CD96 alters the activation, migration, and effector functions of hematopoietic cells, particularly within solid tumors or after specific immune challenges.

  • In vivo depletion or blockade: Clone 3.3 is administered in vivo to achieve functional blockade of CD96, not for cell depletion, but to directly interfere with receptor-ligand interactions, allowing researchers to study immune regulation mechanisms, including possible synergy or antagonism with other immune checkpoints.

Biological context:

  • CD96 (Tactile) is predominantly expressed on T cells and NK cells, making clone 3.3 a specific tool for dissecting their roles in immune homeostasis and disease models, including infection and cancer.

Summary Table: Common In Vivo Applications of Clone 3.3

Application TypePurposeCell Targets
Tumor ImmunologyTest CD96 as an immune checkpoint; boost anti-tumor NK/TNK cells, T cells
Functional BlockadeBlock CD96–ligand interaction in immune studiesNK cells, T cells
Immunoregulation ResearchModulate/explore effector and regulatory immune responsesHematopoietic
Phenotype CharacterizationStudy impact of CD96 on cell activity and migrationNK cells, T cells

All sources agree on the main experimental uses but may differ on model specifics; suppliers and research summaries are most authoritative for antibody applications.

Commonly used antibodies or proteins studied alongside "3.3"—assuming you are referring to 14-3-3 proteins (based on usage in molecular and cell biology literature)—include targets that are known 14-3-3 binding partners, regulatory kinases, or signaling intermediates.

Key co-analyzed proteins/antibodies with 14-3-3 include:

  • Phosphorylated client proteins: Many studies assess the interaction of 14-3-3 with phosphorylated forms of client proteins such as CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) and LRRK2 (Leucine-rich repeat kinase 2) for motif-binding analysis.
  • Raf-1 kinase: A classic binding partner. 14-3-3 forms complexes with phosphorylated Raf-1, which can be assayed using antibodies against Raf-1 or phospho-specific Raf-1 antibodies.
  • Gab2 (Grb2-associated binder 2): Especially in oncology research, Gab2 and its phosphorylated forms are tested in conjunction with 14-3-3, often using anti-Gab2 (total and phospho-specific) antibodies.
  • Tau: In neuroscience, 14-3-3 interaction with phosphorylated tau is commonly analyzed using anti-14-3-3 and anti-phospho-tau antibodies.
  • TASK3 channel: Particularly when studying motif III interactions, as FC-THF and related compounds can alter 14-3-3’s interaction with this potassium channel.

Experimental methods often involve:

  • Immunoprecipitation with anti-14-3-3 and co-staining with antibodies against suspected partner proteins (e.g., phospho-specific antibodies for signaling kinases or channels).
  • Functional assays using small molecule modulators and quantified by immunoblots with antibodies against 14-3-3, its isoforms, and client proteins.

Other antibodies/proteins sometimes included:

  • GSK3 (Glycogen synthase kinase 3)
  • AKT kinase
  • BAD and FOXO (apoptosis and transcription factors)
  • p53 or CDC25 (cell cycle regulators)
  • Isoform-specific antibodies against various 14-3-3 family members (e.g., β, γ, ζ, δ, ε, η, σ, τ).

Alternative 3.3 usage: If you meant antibody isotype IgG3.3 or a different target, please specify, as these are not as universally standardized and co-targets would depend on context.

Summary Table: Proteins/Antibodies Commonly Used with 14-3-3 in Literature

CategoryExample Protein/AntibodyContext
Signaling KinasesRaf-1, LRRK2, AKT, GSK3Signal transduction, phosphorylation sites
Scaffold/AdaptorGab2, IRS-1Complex formation, pathway regulation
Disease MarkersPhospho-Tau, CFTRNeurobiology, cystic fibrosis studies
Cell Cycle/Aptosisp53, CDC25, BAD, FOXOCell fate and survival studies
ChannelsTASK3Ion channel modulation
Isoform-Specific14-3-3 β, γ, ζ, ε, etc.Isoform-dependent signaling

If your context is different (e.g., a specific antibody clone "3.3"), please clarify, but most published data on "3.3" refers to the 14-3-3 protein family.

No search result directly addresses "clone 3.3" citations in the context of code, biology, medicine, or any specific scientific field. The last three search results mention the word "clone," but none provide information about "clone 3.3," nor do they summarize findings from citations of such a clone in the literature.

Your query could refer to a specific software tool, cell line, protein, gene, or another scientific entity—but without clarification or relevant results, it is not possible to summarize key findings from "clone 3.3" citations in the scientific literature based on the provided sources. If you have a specific scientific context or more details about "clone 3.3," please clarify so I can give a more targeted and accurate answer.

Dosing regimens for clone 3.3 (an anti-mouse CD96 monoclonal antibody) vary depending on the mouse model used, experimental goals, and desired immunological outcomes. The specific dose, frequency, and route of administration may be adjusted for differences in mouse strain, disease model, or the intended effect such as cell depletion versus immunomodulation.

  • Dose per Mouse: Reported doses of clone 3.3 often range in the literature from about 200 μg to as high as 500 μg per mouse, depending on the degree of immune intervention required and mouse strain sensitivity.
  • Frequency and Duration: Typical regimens involve administration every 2–3 days (for acute interventions) or weekly for longer-term immunomodulation. The number of doses can further vary with the experimental timeline (for example, repeated for 2–4 weeks in chronic studies).
  • Route of Administration: Clone 3.3 is generally delivered via intraperitoneal (IP) injection for systemic effects, but may also use intravenous (IV) or local routes for specific tissue targeting if required by the experimental design. Variations in absorption and biodistribution can affect efficacy.

Key factors influencing dosing across mouse models:

  • Mouse strain: Sensitivity, immunological baseline, and metabolism can affect the optimal dose. For instance, C57BL/6 versus BALB/c mice may require dosing adjustment based on genetic background or tumor model.
  • Disease model: Tumor-bearing mice might receive more frequent dosing compared to infectious disease or autoimmunity models.
  • Goal of treatment: Cell depletion protocols (e.g., for depleting NK or T cells) may use higher or more frequent doses than protocols aimed at blocking a receptor.

Summary Table: Typical Dosing Regimen Properties for Clone 3.3

Mouse Model/StrainDose per Mouse (μg)RouteFrequencyExperimental Goal
C57BL/6 (tumor model)200–500IPEvery 2–3 daysImmunomodulation, depletion
BALB/c (immunology study)200–300IP/IVWeeklyFunctional blockade
  • Dosing can deviate from these standards based on specific experiment needs, pilot studies, or published protocols for similar clones (see anti-mouse CD96 and other immune cell depleting antibodies).
  • Always adjust regimens based on published guidance, preliminary titrations, and the specifics of the disease/tissue model.

Exact clone 3.3 regimens should be validated for each new experimental setting, taking into account mouse genetics, disease burden, and desired immune impact. For additional details, refer to peer-reviewed antibody dosing guides and product-specific recommendations.

References & Citations

1. Georgiev H, Ravens I, Papadogianni G, et al. Front Immunol. 9:1072. 2018.
2. Chan CJ, Martinet L, Gilfillan S, et al. Nat Immunol. 15(5):431-438. 2014.
3. Roman Aguilera A, Lutzky VP, Mittal D, et al. Oncoimmunology. 7(5):e1424677. 2018.
4. Blake SJ, Stannard K, Liu J, et al. Cancer Discov. 6(4):446-459. 2016.
B
Flow Cytometry
in vivo Protocol

Certificate of Analysis

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Formats Available

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