Anti-Mouse H-2Db (Clone B22/249) – Purified in vivo PLATINUM™ Functional Grade

Clone B22/249 is a monoclonal antibody that specifically targets the mouse H-2Db MHC class I molecule, and its common in vivo applications in mice center on immunological research, particularly for manipulating or studying lymphocyte populations.

Key in vivo applications include:

• Cytotoxicity assays: Used to mediate or assess the specific lysis of target cells expressing H-2Db molecules.
• In vivo cell depletion: Administered to mice to deplete H-2Db-expressing cells, such as CD8+ T cells, by antibody-dependent cellular cytotoxicity (ADCC) or related mechanisms.
• Immunoprecipitation (IP) of H-2Db+ cells or complexes in vivo: Useful in studying antigen presentation, T cell recognition, or characterizing immune cell populations in live animals.
• Functional studies in cancer or infectious disease models: By depleting or functionally blocking certain immune cells, researchers assess the role of H-2Db-restricted immune responses in tumor rejection, viral clearance, or immune regulation.

The antibody is formulated for low endotoxin levels and high purity, ensuring suitability for in vivo administration, minimizing non-specific immune activation.

Summary table of common in vivo applications of B22/249 in mice:

B22/249 is routinely referenced in protocols analyzing antigen presentation, immune cell subset function, and CD8+ T cell responses in murine models.

Commonly used antibodies or proteins alongside B22/249 (which targets mouse H-2Db, a major MHC class I molecule) in the literature include:

• Anti-H-2Kb antibodies (e.g., clone Y3): These target a different MHC class I molecule (H-2Kb) and are frequently used alongside B22/249 to distinguish between the major MHC class I subtypes in mice.
• Monoclonal antibody 20-8-4s: Another commonly used antibody that can immunoprecipitate MHC class I molecules, often included with B22/249 and Y3 in immunoprecipitation protocols.
• Antibodies against CD3/TCR and CD8: These proteins are direct receptors/ligands of MHC class I, and while not always used in the same immunoprecipitation or depletion assays, are relevant in studies involving B22/249 due to their roles in antigen presentation.
• Beta-2 microglobulin (β2m): Not an antibody, but a key protein that forms part of the MHC class I complex; immunoprecipitation methods often assess β2m association with H-2Db using B22/249.
• Flow cytometry and immunoprecipitation panels: Include B22/249 and Y3 as standard controls, sometimes alongside antibodies for other lymphocyte markers.

In summary, anti-H-2Kb (Y3), 20-8-4s, and β2m are the most frequently cited alongside B22/249, especially for distinguishing MHC class I variants and complex analysis in immunology research.

Key findings from scientific literature citing clone B22/249 center on its specificity, functional applications in immunology, and crucial role in characterizing and manipulating the murine MHC class I molecule H-2D^b^. Clone B22/249 is widely used as both an in vivo and in vitro antibody reagent to study antigen presentation, MHC class I peptide content, and cytotoxic T cell responses.

Essential insights and applications from cited literature include:

• Specificity: B22/249 is a murine IgG2a kappa monoclonal antibody that binds specifically to the H-2D^b^ molecule, which is a class I MHC protein found on mouse lymphocytes and involved in antigen presentation to CD8^+^ T cells.

• Application in Immunopeptidomics: The antibody is fundamental for immunoaffinity purification of H-2D^b^-peptide complexes from mouse tissues. Studies have used B22/249 to isolate MHC I–associated peptides for mass spectrometry, mapping the murine MHC class I immunopeptidome across multiple tissues, yielding hundreds of thousands of peptide identifications. This work underpins research into tissue-specific immune presentation and antigen discovery.

• Functional Immunology Studies: B22/249 is often used in experiments evaluating MHC class I function, antigen recognition, and cytotoxic T cell activity. For example, the antibody has been used to:

  • Inhibit or block cytolytic T lymphocyte (CTL) interactions with H-2D^b^, thereby directly demonstrating the molecular requirements for CTL-mediated lysis in vitro.
  • Confirm loss or absence of H-2D^b^ expression in knockout or gene-edited models by flow cytometry and immunofluorescence analyses.

• Proteomics and Quantitative Analysis: B22/249 is featured in protocols that quantify changes in the pool of MHC I–presented peptides, especially in response to therapeutic interventions or cellular stress. For instance, it has been used for isobaric tagging and multiplexed mass spectrometry, enabling precise relative quantitation of low-abundance peptides and investigation of antigen presentation under dynamic biological conditions.

• Transgenic and Knockout Mouse Models: The antibody supports validation and study of mice engineered for H-2D^b^ deficiency or replacement, such as MHC class I knockouts or HLA-A2.1 transgenic mice, making it important for dissecting the role of antigen presentation in immune education and cytolytic activity.

• Alternate Names and Hybridoma Information: In some literature, B22/249 is referred to as B22-249.R1 or H-2.m2; it was generated by immunizing mice with allogeneic spleen cells and is widely distributed through several suppliers for research use.

In summary, clone B22/249 is widely cited as a gold-standard antibody for murine H-2D^b^ detection, purification, and functional blocking, underpinning advances in immunopeptidomics, T cell immunology, and mouse genetics.

Dosing regimens for clone B22/249 (anti-mouse H-2Db) can vary across different mouse models and research applications, but direct, model-specific dosing guidance is not detailed in the main sources found. Instead, most product documentation and related research describe this antibody as a functional-grade reagent for in vivo work, primarily in immunological and cancer studies, but generally do not provide explicit dose, frequency, or route information specific to mouse strain, disease model, or target outcome.

Context and supporting details:

• Clone B22/249 is a monoclonal antibody targeting H-2Db, a major histocompatibility complex (MHC) class I molecule on mouse lymphocytes.
• It is commonly used for in vivo depletion, cytotoxicity assays, and studies involving immune modulation, especially in the context of cancer and immunology research.

General in vivo dosing practices:

• For functional-grade monoclonal antibodies used in mouse models (such as anti-MHC or immune checkpoint antibodies), a common dose range is 100–250 μg per mouse per injection, usually administered via intraperitoneal (i.p.) injection at intervals ranging from every 3 days to once weekly, depending on antibody half-life, application, and desired effect.
• Exact dosing of B22/249 must often be empirically optimized for each study, guided by experimental design and pilot studies, as suppliers and protocols frequently do not specify a single recommended regimen for all models.

Key considerations influencing dosing regimens:

• Mouse strain (e.g., C57BL/6, BALB/c) and health status
• Disease model (e.g., tumor type, infection, autoimmune disease)
• Desired biological outcome (e.g., cell depletion, blockade, or labeling)
• Route of administration—i.p., intravenous (i.v.), or local injection
• Frequency and duration required for sustained target engagement or depletion

Relevant product information:

• B22/249 is often supplied at concentrations ≥5.0 mg/ml and in sizes ranging from 1 mg to 100 mg.
• Low endotoxin and high purity formulations make it suitable and safe for repeated in vivo administration.
• Storage and handling protocols are standardized (short-term at 2–8°C, long-term at −70°C).

Summary Table (General dosing reference for in vivo mouse antibodies):

No published data was identified specifying unique dosing schedules of B22/249 based on particular mouse models (like specific strains or disease backgrounds). For high experimental accuracy, pilot studies or dose titration based on response/readout and consulting supplier technical support are recommended when using B22/249 in a new model.

If a published study using B22/249 in a specific mouse model is required, further targeted literature review may be necessary, as most product and protocol information remains general and does not differentiate between models.