Anti-Mouse H-2K^d (MHC Class I) [Clone A4C8.1-Do9] — Purified in vivo GOLD™ Functional Grade

Clone A4C8.1-Do9 is used in in vivo mouse studies as a monoclonal antibody that specifically targets MHC Class I H-2Kd molecules on mouse cells.

This antibody is utilized primarily to:

• Bind to and block MHC Class I H-2Kd molecules: This can be used for functional studies of immune modulation, such as depleting CD8+ T cell responses, interfering with antigen presentation, or investigating the role of MHC Class I in disease models.
• Facilitate transplantation, autoimmune, or immunology experiments: By blocking or neutralizing MHC Class I, researchers can analyze how disruption of these molecules influences immune rejection, tolerance, or infection.

Key features:

• Applications: While often used in western blotting (WB), the "In Vivo" label and low endotoxin preparation make it suitable for injection into live mice, minimizing inflammatory side effects.
• Specificity: Recognizes H-2Kd, which is expressed on cells from mice with the appropriate haplotype (excluding red blood cells); relevant for strains such as BALB/c.
• Usage notes: Each investigator must optimize the dosing and schedule for their experimental needs.

Summary table:

These antibodies are strictly for research use only and not for diagnostic or therapeutic applications. The exact protocol (e.g., dose, route, timing) varies according to experimental design, so investigators need to calibrate usage according to their model requirements.

A4C8.1-Do9 is a monoclonal antibody, though the search results do not specify its target or application, so this answer is based on standard antibody co-utilization practices in immunology, especially as suggested by the provided literature.

In studies utilizing antibodies like A4C8.1-Do9, researchers commonly use the following antibodies and proteins alongside it to investigate immune responses, tissue environments, or co-localized antigens:

• F4/80: A marker for macrophages, often co-stained to assess immune cell infiltrates in tissues.
• Ly6C: Used to distinguish monocyte subpopulations in tissue or circulating blood.
• IL-1β: Important cytokine, often stained to assess inflammatory responses and cellular activation.
• XCR1: A marker for a subset of dendritic cells involved in antigen presentation, often visualized to study immune cell clustering.
• CD8 and CD4: For identifying cytotoxic or helper T cell subsets within immune responses.
• Antibodies against AAV capsid or transgene product: Often used in gene therapy or immunology research to monitor antigen-specific T cell responses.
• Secondary antibodies: Species-specific, conjugated with fluorescent dyes or enzymes for signal detection in immunofluorescence (IF) or immunohistochemistry (IHC).
• Other cell markers: Such as CD40/CD40L (co-stimulatory interactions), MyD88 (innate immune signaling), and various cytokines for the functional study of cell pathways.

These antibodies are typically chosen to allow multiplex detection of different cell types or proteins:

• By selecting antibodies from different host species (e.g., mouse, rabbit), researchers avoid cross-reactivity when using multiple monoclonals in the same experiment.
• Using distinct fluorochromes or chromogenic substrates ensures clear, simultaneous visualization of target antigens.
• Controls such as isotype-matched antibodies also play an essential role in validating specificity.

The co-use of F4/80, Ly6C, IL-1β, XCR1, and CD8/CD4 antibodies is particularly common in studies analyzing immune cell dynamics in tissues—such as liver following gene therapy protocols—where cellular infiltration, activation, and co-localization are critical outcomes.

If a specific protein or antigen context for A4C8.1-Do9 is needed, providing its target or application would allow for a more precise answer. Otherwise, these listed antibodies represent the standard panel of immune markers often used in tandem with application-specific monoclonals in immunological and tissue-based research.

Clone A4C8.1-Do9 is a monoclonal antibody that specifically recognizes the major histocompatibility complex (MHC) class I H-2K^d haplotype in mice, which is ubiquitously expressed on nucleated cells of this genotype.

Key findings and characteristics from scientific citations include:

• Specificity: Clone A4C8.1-Do9 is specific for an epitope on mouse MHC class I H-2K^d, distinguishing cells from strains such as BALB/c (H-2K^d haplotype), and is used to identify and characterize these cells in immunological studies.
• Applications: It is widely used in immunological assays including flow cytometry, ELISA, and Western blotting to label and quantify cells expressing H-2K^d.
• Antibody Details:
  • Monoclonal, mouse origin, isotype IgG1.
  • Available conjugated to allophycocyanin (APC) or as a low-endotoxin, functional-grade antibody for in vivo and in vitro applications.
  • Typical concentration for staining cells in flow cytometry is ~0.25 µg per 10^6 cells in 100 µl.
• Handling: Stable for short-term storage at 2–8°C; for long-term, aliquot and store at -80°C to avoid freeze-thaw cycles.
• Biological Context: MHC class I molecules such as H-2K^d mediate antigen presentation to cytotoxic T cells and are a critical target for immunological research involving transplant models, immunophenotyping, and studies of immune cell activation.
• Research Utility: Clone A4C8.1-Do9 provides a reliable tool for distinguishing between mouse strains based on MHC haplotypes, tracking donor cell populations, and studying immune responses in experimental models.

No significant controversies or conflicting results are reported in the cited literature. Clone A4C8.1-Do9 is recognized as a standard reagent in murine immunology research for analysis of H-2K^d expression.

Dosing regimens for clone A4C8.1-Do9 primarily depend on the application and mouse model, but available guidance is specific for in vitro use, especially flow cytometry: the recommended concentration is ~0.25 µg per 10^6 cells in a 100 µl volume.

Most published information relates to use for cell staining in flow cytometry rather than in vivo experiments. For flow applications (all mouse models):

• Concentration: ~0.25 µg per 10^6 cells in 100 µl.
• Optimal titration: Titration is recommended to tailor the dose to specific cells, strains, or experimental setups.

For in vivo dosing (e.g., systemic administration or functional studies), no standard regimen for A4C8.1-Do9 is available in the search results as of the latest guidance. For other functional-grade antibodies (used for depletion or functional blockade), doses typically range from 100–300 µg per mouse per dose given intraperitoneally, 1–3 times per week, depending on the target and the effect sought. However, this should not be extrapolated to A4C8.1-Do9 without further validation, as the antibody is reported only for in vitro applications.

Key considerations for mouse model differences:

• The source does not explicitly differentiate dosing of A4C8.1-Do9 across mouse strains, disease models, or genetic backgrounds.
• Investigators are advised to empirically titrate the antibody for each application and potentially each mouse model.

If using A4C8.1-Do9 for functional in vivo studies, it is essential to consult lot-specific datasheets and titrate accordingly, as the manufacturer notes that each investigator must determine the optimal working dilution for their specific context.

Summary Table: A4C8.1-Do9 Dosing Guidance

No specific regimen differing by mouse model is documented; empirical titration remains the standard approach for optimizing use of this clone in different systems.