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 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.

Dosing regimens for clone B22/249 anti-mouse H-2Db antibodies are not universally standardized and tend to vary depending on the specific mouse model, experimental goals (such as depletion vs. functional blockade), and the institution's protocols. However, direct published data describing specific dosing regimens for B22/249 across diverse mouse models are limited in the available literature.

Key details from available sources:

• B22/249 Properties: This antibody is offered as a functional-grade reagent for in vivo use, with concentrations typically provided at ?5.0 mg/ml and available in aliquots ranging from 1 mg to 100 mg. The product is used for cytotoxicity assays, in vitro depletion, and immunoprecipitation.
• Guidance for Mouse Antibody Dosing: For comparable in vivo monoclonal antibody targeting major histocompatibility complex (MHC) molecules or immune checkpoint molecules in mice, typical doses range from 100–250 ?g per mouse, administered by intraperitoneal injection every 3–4 days, as seen with anti-PD-1 or anti-CTLA-4 clones. While anti-H-2Db is not specifically listed, the dosing principles are similar for depleting or blocking monoclonal antibodies in immunological mouse models.
• Application Context: Dosing may be tailored in syngeneic tumor models, depletion protocols, or transplantation studies, with adjustments according to the desired degree of cellular depletion or blockade and the pharmacokinetics in the specific mouse strain (e.g., C57BL/6 used as a standard background).
• Published Example with B22/249: A study using purified B22/249 provides indirect support for these standard dosing philosophies but does not specify the microgram or milligram amount used per mouse.

Summary Table: Typical In Vivo Dosing for Mouse Monoclonal Antibodies
| Antibody (example clone) | Standard Dose per Mouse | Frequency | Route | Application ||-------------------------------|------------------------|--------------------|--------------------|------------------|| B22/249 (anti-H-2Db) | Not explicitly stated* | Likely every 3–4d | Intraperitoneal** | Depletion/blockade|| RMP1-14 (anti-PD-1) | 200–500 ?g | Every 3–4d | Intraperitoneal | Checkpoint blockade|| 30F11 (anti-CD45) | 100–250 ?g | Every 3–4d | Intraperitoneal | Depletion |

* Exact dose for B22/249 is not stated in available sources but is likely analogous to other functional IgG2a mouse monoclonal antibodies (100–250 ?g per mouse).
** Most functional delivery is via i.p. injection for systemic effect.

Additional notes:

• Always tailor dosing for B22/249 to the specific experimental model, considering strain, age, sex, administration route, and endpoint (blockade vs. cell depletion).
• If using for depletion, pilot studies may be required to titrate the lowest dose that yields efficient depletion without toxicity.

If you require precise, peer-reviewed doses for a particular disease model or protocol, these are often specified in the Materials and Methods sections of primary research articles, or can be provided by the antibody supplier upon request.