Anti-Mouse H-2Kb (MHC Class I) [Clone Y-3] – Purified in vivo GOLD™ Functional Grade

Clone Y-3 is most commonly used in vivo in mice to block MHC class I (specifically H-2K^b^ and related haplotypes) function, thereby inhibiting MHC class I–mediated T cell responses.

Key in vivo applications include:

• MHC Class I Blockade: Clone Y-3 is frequently employed to transiently block MHC class I molecules (notably H-2K^b^) in mice, allowing researchers to study the effects of inhibiting CD8^+^ T cell responses. This is important for dissecting the role of MHC class I–restricted antigen presentation in immunity, autoimmunity, transplantation, and tumor rejection models.
• Functional Immunological Assays: By blocking MHC class I, clone Y-3 is used to test the dependence of various immune responses on MHC class I-mediated antigen presentation. These include responses to infections, graft-versus-host disease (GVHD), and tumor immunity.
• In Vivo Depletion/Functional Inhibition: Administration of clone Y-3 can functionally disable MHC class I on the cell surface in vivo, which helps assess the consequences of absent CD8^+^ T cell recognition in different experimental contexts.

Secondary laboratory applications (in vitro and ex vivo, but sometimes related to in vivo studies) include:

• Flow Cytometry: Detecting H-2K^b^ expression levels on cells from treated or experimental animals.
• Immunoprecipitation and Cytochemistry: Analysis of MHC class I expression as part of tissue or cell characterization efforts following in vivo interventions.

Notes:

• Specificity: Y-3 binds to H-2K^b^, H-2K^k^, H-2K^q^, H-2K^r^, and H-2K^s^, but not H-2K^d^, so choice of mouse strain is crucial.
• The use of this clone enables researchers to directly probe the consequences of blocking antigen presentation to CD8^+^ T cells in living mice, supporting studies in cancer immunology, viral pathogenesis, and transplantation immunology.

There is some confusion in the literature with clone Y-3 versus Y-3P; Y-3P is an anti–MHC class II (I-A) antibody, while Y-3 is specific for MHC class I (H-2K^b^). This answer pertains to clone Y-3 (not Y-3P) and its in vivo uses.

Other commonly used antibodies or proteins applied in conjunction with Y-3 (anti-mouse H-2K^b, MHC Class I, Clone Y3) in the literature include actin, GFP, CD markers such as CD31, and various secondary antibodies for detection. These are employed to assess protein loading, label specific cell types, or facilitate visualization and quantification in immunological and cell biology assays.

Key details:

• Actin: Frequently used as a loading control in western blotting and immunostaining to confirm equal protein input.
• GFP (Green Fluorescent Protein): Used for identifying transgenic populations or reporter expression in conjunction with Y-3 to phenotype or gate specific cells.
• CD markers (e.g., CD31): Utilized to label and distinguish immune or endothelial cell populations in multi-parameter flow cytometry or imaging studies.
• Secondary antibodies: Applied for detection when the Y-3 antibody is not directly conjugated to a detection moiety (e.g., fluorophore or enzyme).

Other studies may include antibodies against other MHC class I or II proteins and proteins relevant to the experimental disease model or transgenic strain under investigation. Frequently, multi-antibody panels are built to dissect complex cellular landscapes or monitor immune cell activation and trafficking.

In summary, Y-3 is most commonly paired with structural proteins (like actin), lineage or state markers (CD markers, GFP), and secondary detection reagents. This enhances cell-type specificity and methodological rigor across diverse immunological assays.

Key Findings from clone Y-3 (Anti-H-2Kb) in Scientific Literature

Specificity and Application

• Clone Y-3 is a monoclonal antibody that specifically identifies and binds mouse H-2Kb molecules, a major histocompatibility complex (MHC) class I antigen.
• It is commonly used to block H-2Kb function in vivo and is also validated for techniques such as Western blotting, immunoprecipitation, immunocytochemistry, flow cytometry, and functional assays.
• The antibody is recognized for its use in identifying and modulating MHC class I interactions, which are central to immune responses including antigen presentation and T cell recognition.

Functional Insights

• In experimental settings, clone Y-3 is employed to block or knock out the function of H-2Kb, allowing researchers to study the consequences of MHC class I molecule deprivation on immune cell behavior or immune responses.
• Functional blockade with clone Y-3 helps elucidate the role of H-2Kb in immune tolerance, T cell activation, and evasion of immune surveillance, particularly in model systems like cancer and autoimmune research.
• The clone is widely cited for its specificity, reliability, and utility in mechanistic studies involving MHC class I molecules in mice.

Availability and Validation

• Clone Y-3 is available from several suppliers and consistently advertised as “functional grade,” indicating it is suitable for in vivo and in vitro applications.
• The antibody has been validated in multiple assay formats, reflecting its versatility and widespread adoption in immunological research.

Limitations in Literature Coverage

• No direct citations to primary scientific literature or detailed mechanistic studies referencing clone Y-3 appear in the available results from major repositories or through summaries from manufacturers.
• The majority of “key findings” are summarized by reagent providers and not by peer-reviewed journal articles, suggesting that while clone Y-3 is widely used, its specific applications and findings may be dispersed across a range of studies and not always highlighted in abstracts or citation summaries.
• This lack of centralized citation information suggests researchers should look to protocols and methods sections of relevant immunology papers for detailed use cases and experimental outcomes involving clone Y-3.

Summary Table

Conclusion

Clone Y-3 is a well-established tool for identifying and functionally blocking mouse H-2Kb, with recognized specificity and utility across a range of immunological assays. While it is widely referenced and available commercially, detailed mechanistic findings or breakthrough discoveries specifically attributed to clone Y-3 in the scientific literature are scarce in the context of citation summaries; most references are from reagent providers summarizing its general use and validation. Researchers should consult primary literature and experimental methods for in-depth examples of clone Y-3 in action.

The dosing regimens for clone Y-3, an antibody targeting mouse MHC Class I (H-2Kb), can vary significantly across different mouse models. While specific dosing protocols for clone Y-3 are not detailed in the current search results, general guidelines for similar antibodies suggest that dosing often depends on several factors:

1. Mouse Strain: Different mouse strains may require adjustments due to genetic variations that could affect immune responses or the expression of targeted epitopes.

2. Experimental Context: The specific experimental conditions, such as the goal of the study (e.g., immune modulation, cell depletion), can influence the chosen dosing regimen.

3. Route of Administration: Common routes include intraperitoneal (i.p.) or intravenous (i.v.) injections, with each route potentially impacting the efficacy and distribution of the antibody.

4. Frequency of Administration: Dosing schedules can range from once every few days to more frequent injections, depending on the study's objectives and the dynamics of the immune system in the model.

For antibodies targeting similar immune components, dosages often range from 200 to 500 μg per mouse per injection, with administration every 2 to 3 days being common. However, without specific studies or protocols for clone Y-3, these general guidelines from related antibodies should be considered as a starting point, and actual dosing may need to be optimized based on empirical data from specific mouse models.