Anti-Mouse H-2K^d (MHC Class I) – Purified in vivo PLATINUM™ Functional Grade

Clone A4C8.1-Do9 is a monoclonal antibody that recognizes mouse MHC class I H-2K^d and serves multiple important functions in mouse immunological research. This antibody has several well-established in vivo applications that leverage its specificity for the H-2K^d haplotype.

Immune System Studies

The primary in vivo applications of clone A4C8.1-Do9 involve investigating immune responses and immunological processes in mice. Researchers commonly employ this antibody in studies examining T cell function, as MHC class I molecules are critical for presenting antigens to CD8+ T cells and initiating cellular immune responses.

Transplantation Research

Clone A4C8.1-Do9 is frequently used in transplant rejection studies. Since MHC class I molecules play a central role in allograft recognition and rejection, this antibody enables researchers to track donor cell populations and monitor immune responses following transplantation procedures. The antibody's ability to distinguish between mouse strains based on MHC haplotypes makes it particularly valuable for these applications.

Strain Identification and Cell Tracking

A key utility of this clone is its capacity to differentiate between mouse strains that express different MHC haplotypes. This makes it an essential tool for tracking donor cell populations in experimental models, particularly in mixed chimera studies or adoptive transfer experiments where distinguishing between host and donor cells is critical.

Technical Considerations

For in vivo applications, the antibody is available in functional grade formulations with low or ultra-low endotoxin levels (≤1.0 EU/mg). These preparations are manufactured using animal-free, protein-free cell culture techniques and undergo rigorous purification to minimize endotoxins and aggregates, ensuring they are suitable for administration to live animals without triggering unwanted inflammatory responses.

Commonly used antibodies and proteins in combination with A4C8.1-Do9 (an anti-MHC class I H-2K^d antibody) in the literature include markers and antibodies that characterize various immune cell populations and functional states.

Frequently co-used antibodies or proteins include:

• F4/80 (macrophage marker)
• Ly6C (monocyte and neutrophil differentiation)
• IL-1β (pro-inflammatory cytokine)
• XCR1 (dendritic cell marker)
• CD8 and CD4 (T cell subset markers)

These antibodies are used to delineate different immune cell types within tissues (such as liver, spleen, or sites of inflammation), often in the context of immunophenotyping by flow cytometry or immunohistochemistry. In studies involving immune response, transplant rejection, or T cell function, A4C8.1-Do9 is often part of a panel that includes other major immune markers for comprehensive cellular analysis.

Additionally, in general immunodetection workflows, secondary antibodies (such as anti-mouse IgG conjugated to fluorophores or enzymes) are used to visualize or quantify primary antibodies like A4C8.1-Do9 via techniques such as flow cytometry, immunofluorescence, or western blot.

To summarize, A4C8.1-Do9 is most commonly paired with:

• Immune cell subset markers (F4/80, Ly6C, CD4, CD8)
• Cytokines (IL-1β)
• Dendritic and myeloid cell markers (XCR1)
• Appropriate secondary antibodies or detection reagents depending on assay type.

These combinations allow for precise identification and functional analysis of cell populations in murine immune research.

Clone A4C8.1-Do9 is a monoclonal antibody that has been extensively used in immunological research for its highly specific recognition of the mouse MHC class I H-2K^d haplotype. The key findings from scientific literature regarding this clone reveal its important characteristics and applications in cellular immunology.

Specificity and Target Recognition

Clone A4C8.1-Do9 recognizes a specific epitope on mouse MHC class I H-2K haplotype d (H-2K^d). This specificity makes it particularly valuable for distinguishing cells from mouse strains carrying the H-2K^d haplotype, such as BALB/c mice, and is used to identify and characterize these cells in immunological studies. The antibody targets the major histocompatibility complex (MHC) class I molecule, which is ubiquitously expressed on the cell surface of nucleated cells.

Structural Context

The MHC class I molecule recognized by this clone consists of a 45-kDa type I transmembrane glycoprotein (α-chain or heavy chain) and a 12-kDa soluble protein (β2-microglobulin, β2M). The α-chain comprises three domains (α1, α2, and α3), where α1 and α2 form the closed antigen-binding groove that binds to 8-10 amino acid peptides derived from cytosolic antigens. The β2M noncovalently associates with the α3 domain, which is essential for MHC stability.

Applications in Flow Cytometry

The antibody has been optimized for flow cytometry applications, with recommended concentrations of ≤0.25 μg per 10^6 cells in a volume of 100 μl or 100 μl of whole blood. Titration of the reagent is recommended for optimal performance for each specific application.

Based on the available information, there is no standard dosing regimen for clone A4C8.1-Do9 across different mouse models, as dosing primarily depends on the specific application rather than broad variability across mouse strains.

Flow Cytometry Applications

For flow cytometry applications, clone A4C8.1-Do9 antibody has a well-defined concentration guideline. The suggested concentration is ≤0.25 μg per 10⁶ cells in a volume of 100 μl or 100 μl of whole blood. However, titration of the reagent is recommended for optimal performance in each specific application.

Western Blot and Other Applications

For Western blot applications, investigators must determine their own optimal working dilution, as requirements vary based on specific experimental conditions. Directions may also vary by lot and should be consulted on lot-specific datasheets, as information may periodically change.

Limited In Vivo Dosing Information

Unlike other commonly used antibodies such as anti-CTLA-4 (9H10, 9D9), anti-CD4 (GK1.5), or anti-CD8 (2.43), which have established standard dose ranges of 100-250 μg per mouse for in vivo studies, specific in vivo dosing protocols for clone A4C8.1-Do9 are not available in the current literature. This antibody, which recognizes the MHC class I H-2K haplotype d (H-2Kd), appears to be primarily utilized for cell staining and analytical purposes rather than therapeutic interventions in mouse models.

Researchers planning to use this antibody should consult manufacturer guidelines and conduct preliminary dose-finding studies tailored to their specific experimental design and application.