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

Clone B22/249 is a monoclonal antibody targeting mouse H-2Db, an MHC class I molecule, and is used in in vivo mouse studies primarily for immunological research applications such as cytotoxicity assays, in vivo cell depletion, and immunoprecipitation.

Key usage contexts:

• Functional in vivo studies: B22/249 is formulated as a functional-grade, low-endotoxin antibody specifically suitable for direct administration in live mice. Its main research applications include:
  • Cytotoxicity assays: Assessing immune cell function and MHC-I–dependent cytotoxic reactions.
  • In vivo depletion: Eliminating or modulating populations of cells expressing H-2Db (such as specific lymphocytes) by antibody-mediated mechanisms; this enables studies of immune system function by selective cell removal.
  • Immunoprecipitation (IP): Isolating MHC-I complexes or associated proteins for downstream analysis.
• Cancer and immunology research: Frequently employed in tumor immunology to investigate antigen presentation, T cell recognition, and mechanisms of tumor immune evasion.
• Antigen specificity: B22/249 binds to H-2Db expressed on the surface of mouse lymphocytes and can be used to identify or manipulate MHC class I–restricted immune responses.

Important technical details:

• Isotype: Mouse IgG2a kappa.
• Purity/endotoxin: Typically prepared at >95% purity and ?1.0 EU/mg endotoxin to ensure safety and experimental reliability in vivo.
• Recommended storage: Sterile, at 2–8°C (short-term); aliquoted and frozen for long-term storage.

In summary, clone B22/249 is a well-established tool for modulating and analyzing mouse MHC class I–mediated immune responses in vivo, including cell depletion and cytotoxicity studies, with particular relevance in preclinical cancer and immunology models.

The correct storage temperature for sterile packaged clone B22/249 (anti-mouse H-2Db antibody) depends on the intended duration of storage:

• Short-term storage: Store at 2–8°C (refrigerator temperature) for up to one month.
• Long-term storage: Store at -20°C, avoiding repeated freeze/thaw cycles.

For optimal preservation, always keep the antibody in its original sterile packaging, minimize light exposure if the antibody is conjugated to a fluorochrome, and aliquot before freezing to prevent activity loss due to freeze/thaw cycles.

Most manufacturers indicate that storage at 2–8°C is suitable for a few weeks to a month, while -20°C is preferred for longer-term stability. Storing at room temperature is not recommended as it leads to degradation.

The B22/249 antibody is commonly used to target the mouse MHC class I molecule H-2Db, particularly in immunology research involving antigen presentation, cytotoxicity assays, and in vivo depletion. In the literature, B22/249 is often used with a set of other antibodies or proteins to address related questions about immune cell function, phenotype, or antigen specificity.

Other commonly used antibodies/proteins with B22/249 include:

• Anti-H-2Kb antibodies (targeting a different mouse MHC class I molecule), often used together with H-2Db antibodies to distinguish between MHC class I subtypes on murine cells.
• CD8 and CD3 antibodies, for T cell identification in flow cytometry or cytotoxic T lymphocyte (CTL) assays; these are standard for immune cell phenotyping when investigating antigen presentation and CTL responses.
• CD4 antibodies, when studies involve helper T cell populations for broader lymphocyte profiling.
• Beta-2 microglobulin antibodies, sometimes used to evaluate proper assembly or expression of MHC class I complexes.
• Isotype-matched control antibodies (e.g., mouse IgG2a), routinely included for gating and specificity controls in flow cytometry and immunoprecipitation.
• Conjugated antibodies (e.g., FITC, PE, biotin)—antibodies may be labeled to allow for multiplexed detection alongside B22/249.

Other relevant clones and closely related antibodies:

• Clone 28-14-8S (anti-H-2Db), used in similar applications as B22/249 and sometimes in tandem in comparative or validation studies.

In summary, when B22/249 is used in the literature, it is commonly paired with antibodies against other MHC class I/II molecules (like H-2Kb), T cell markers (CD3, CD8, CD4), and isotype controls, depending on the experimental context. This allows researchers to comprehensively profile antigen presentation and immune cell function.

Clone B22/249 is a monoclonal antibody specific for the mouse H-2D^b MHC class I molecule, widely used in immunology research. Key findings from scientific literature citing clone B22/249 include the following:

• Specificity and Function: B22/249 is a functional-grade murine IgG2a kappa antibody that binds specifically to the H-2D^b molecule, a class I MHC protein expressed on the surface of lymphocytes in mice. It is frequently used for cell surface labeling, in vitro depletion, and cytotoxicity assays to investigate immune responses, especially those involving CD8⁺ T cells.

• Experimental Use in Immunology: In experiments analyzing antigen presentation and T cell responses, B22/249 is employed to block or deplete H-2D^b-expressing cells. For example, in studies of T cell recognition following immunization, B22/249 is used to pre-incubate stimulator cells to specifically block the presentation of antigenic peptides on H-2D^b and thus confirm the MHC restriction of CD8⁺ T cell responses. This supports precise dissection of cellular immune mechanisms, especially in the context of transplantation, tumor immunology, and infectious disease research.

• Product and Quality: The antibody is formulated for in vivo and in vitro use, with high purity (?95% by SDS-PAGE and SEC), low endotoxin (<1.0 EU/mg), and manufactured under animal-free conditions (via cell culture).

• Reported Applications:

  • Immunoprecipitation (IP)
  • In vivo depletion and targeting
  • Flow cytometry for cell surface detection
  • Functional blocking of MHC class I-mediated antigen presentation

• Research Focus: Citations often reference B22/249 in studies examining transplant rejection, anti-tumor immune responses, and the biology of MHC class I molecules. The antibody aids in elucidating the role of H-2D^b in immune recognition and T cell activation.

• Distribution and Access: B22/249 is available from scientific suppliers (e.g., Cedarlane, Assay Genie) for research use, confirming its broad utility and validation in peer-reviewed studies.

In summary, B22/249 is a critical research tool for blocking or detecting mouse H-2D^b MHC class I molecules, enabling detailed mechanistic studies of CD8⁺ T cell responses in preclinical immunology models.