Anti-Mouse CD90.2 (Thy 1.2) [Clone 30-H12] — Purified in vivo GOLD™ Functional Grade

Common In Vivo Applications of Clone 30-H12 in Mice

Clone 30-H12 is a monoclonal antibody specific for mouse CD90.2 (Thy-1.2) and is widely utilized in various experimental contexts in mice.

T Lymphocyte Depletion

One of the most prominent in vivo uses of 30-H12 is the selective depletion of T lymphocytes in mouse models expressing the Thy1.2 alloantigen (such as C57BL/6, BALB/c, CBA, and several other strains). This application is crucial for studies investigating the role of T cells in immune responses, transplantation, autoimmunity, and cancer. For example, administration of 30-H12 can prevent acute graft-versus-host disease by depleting donor T cells in transplantation models. The antibody is effective at depleting both thymocytes and mature peripheral T cells.

Hematopoietic Stem Cell and Neuron Identification

Beyond depletion, 30-H12 serves as a cell surface marker for identifying specific cell populations, including hematopoietic stem cells and neurons. While this is more commonly an in vitro or ex vivo application (e.g., flow cytometry, immunohistochemistry), the antibody’s specificity makes it valuable for lineage tracing and cell sorting in studies of neurobiology and hematopoiesis.

Functional Studies in Immunology and Neuroscience

In addition to depletion and identification, 30-H12 has been reported to induce calcium flux in thymocytes, suggesting a functional impact on T cell signaling. Its reactivity with neurons also positions it as a tool in neurobiological research, though most neuron-targeted applications are performed ex vivo. The antibody’s ability to modulate T cell function makes it useful for dissecting mechanisms underlying immune tolerance, antigen presentation, and lymphocyte activation.

Model Suitability

It is important to note that clone 30-H12 does not react with the Thy-1.1 alloantigen, so it is not suitable for mouse strains such as AKR/J and PL that express Thy-1.1 instead of Thy-1.2. Successful application therefore requires confirmation of the mouse strain’s Thy-1 allotype.

Summary Table: Major In Vivo Uses of Clone 30-H12

Key Points

• Clone 30-H12 is primarily used in vivo for T lymphocyte depletion in Thy1.2+ mouse strains, enabling studies of immune function, transplantation, and autoimmunity.
• It is a critical tool for preventing graft-versus-host disease in mouse models.
• The antibody can induce functional changes in T cells, such as calcium flux.
• It is also a marker for specific cell types, though most identification applications are performed ex vivo.
• Strain specificity is essential—30-H12 does not recognize Thy-1.1, so it is ineffective in AKR/J and PL strains.

These applications make clone 30-H12 a cornerstone reagent in mouse immunology and, to a lesser extent, neurobiology research.

Commonly Used Antibodies and Proteins with 30-H12

The 30-H12 monoclonal antibody targets CD90.2 (Thy-1.2) in mice and is widely used in immunophenotyping, flow cytometry, immunohistochemistry, and cell depletion studies. Several other antibodies and proteins are commonly used in combination with 30-H12 in the literature and laboratory workflows.

Frequently Paired Antibodies in Immunophenotyping

In cytometry and cell surface marker profiling, 30-H12 is most commonly used alongside antibodies against:

• CD3: A pan-T cell marker; co-staining with 30-H12 helps distinguish T cell subsets.
• CD4: Identifies helper T cells.
• CD8: Identifies cytotoxic T cells.
• CD25: Marks activated T cells and regulatory T cells (Tregs).
• CD45: A pan-leukocyte marker; helps gate on immune cells.
• Fc receptor blockers: Used to minimize non-specific antibody binding, improving specificity in multicolor flow cytometry.

These combinations are standard in mouse immunology to delineate T cell populations, assess activation status, and characterize immune cell subsets.

Additional Pairings in Functional and Tissue Studies

Beyond classic immune markers, 30-H12 is also used with:

• Epithelial markers: For studies involving epithelial tissues, especially in co-staining experiments to distinguish epithelial from stromal or immune components.
• Neural markers: Since CD90.2 is expressed in neurons and neural tissues, 30-H12 is often paired with neural-specific antibodies in neuroscience research.

Functional Assays and Depletion Studies

In functional assays (e.g., T cell depletion, costimulation, apoptosis), 30-H12 may be used in combination with:

• Depleting antibodies: For targeted T cell removal in vivo or in vitro.
• Costimulatory molecules: To study T cell activation and signaling pathways.
• Apoptosis markers: To investigate cell death in primary T cells or thymocytes.

Notes on Specificity and Cross-Reactivity

The 30-H12 antibody specifically recognizes the Thy-1.2 alloantigen and does not cross-react with strains bearing the Thy-1.1 alloantigen (e.g., AKR/J, PL). It is therefore essential to confirm the mouse strain before use.

Summary Table

Key Points

• 30-H12 is routinely paired with CD3, CD4, CD8, CD25, CD45, and Fc receptor blockers for comprehensive immunophenotyping in mice.
• In tissue studies, it is used with epithelial and neural markers to differentiate cell types.
• Functional studies may combine 30-H12 with depleting antibodies, costimulatory molecules, or apoptosis markers to investigate T cell biology.
• Strain specificity is critical, as 30-H12 does not recognize Thy-1.1-bearing strains.

These combinations reflect the diverse applications of 30-H12 in both basic and translational immunology research.

Clone 30-H12 is a well-characterized monoclonal antibody that specifically binds to mouse CD90.2 (Thy-1.2), and its citations in scientific literature reveal several key findings and broad applications related to T cell biology, immune cell depletion, and signal transduction.

Key findings from scientific citations include:

• Specificity and Expression Patterns:

  • 30-H12 binds specifically to the CD90.2 (Thy-1.2) alloantigen present on thymocytes, most peripheral T lymphocytes, some intraepithelial T lymphocytes, epithelial cells, fibroblasts, neurons, and hematopoietic stem cells in most mouse strains, but does not cross-react with CD90.1/Thy-1.1 or with rat Thy-1.
  • CD90.2 is a GPI-anchored membrane glycoprotein involved in cell–cell interactions and is especially important in the mouse immune system and nervous system.

• Biological Functions Uncovered:

  • CD90.2 interacts with CD45 in signal transduction pathways and plays a role in mediating the adhesion of thymocytes to thymic stroma.
  • Crosslinking 30-H12 antibody on thymocytes can induce Ca²⁺ influx, intensify CD3/TCR-mediated signal transduction, promote apoptosis of thymocytes, and inhibit proliferative responses in mature T cells, highlighting its role in modulating T cell activation and signaling.
  • Evidence suggests an involvement in broader cell-ligand interactions, potentially influencing synaptogenesis and other neural events.

• Experimental and Therapeutic Applications:

  • 30-H12 is widely used for flow cytometry, immunohistochemistry, and in vivo/in vitro depletion of T lymphocytes.
  • Depletion of Thy-1.2⁺ cells using 30-H12 in vivo is a standard technique for investigating the role of T cells in animal models. For example, in a tuberculosis study, depletion of Thy-1.2⁺ cells in vaccinated, CD4-deficient mice abrogated protective immunity, directly establishing the essential contribution of Thy-1.2⁺ T cells to protective responses.
  • 30-H12 is also reported in use for studying myocarditis and evaluating immune cell contributions in various disease models.

• Technical Properties:

  • Frequently employed as a lineage marker for T cell identification, quantification, and depletion in mice.
  • It does not cross-react with other CD90 alloforms, ensuring high selectivity for Thy-1.2⁺ cells.

Summary Table: Key Aspects of 30-H12 Citations

These findings highlight the importance of clone 30-H12 both as a tool for immune cell characterization/depletion and in illuminating mechanisms of T cell biology and function in mouse models.

Dosing regimens for the clone 30-H12 monoclonal antibody, typically used to target mouse CD90.2 (Thy1.2), can vary depending on the specific application and mouse model used. Here are some general considerations:

1. Depletion of T Lymphocytes

• Dose Range: Generally, for in vivo depletion studies, dosing may range from 100 to 500 μg per mouse. However, specific dosing should be tailored based on the desired level of depletion and the individual study's requirements.
• Route of Administration: Intraperitoneal (IP) injection is commonly used for antibody administration.

2. Immunological Studies

• Flow Cytometry: For flow cytometric analysis, the 30-H12 antibody can be used at concentrations less than or equal to 0.125 μg per test .
• Other Applications: For immunohistochemistry or immunoprecipitation, the optimal dose may vary based on the specific protocol and tissue being studied.

3. General Considerations

• Mouse Strains: The effectiveness of the 30-H12 antibody can vary among mouse strains, as it specifically targets Thy1.2, which is not present in all strains .
• Antibody Formulation: Ensure that the antibody is of functional grade (such as purified form) when used for in vivo applications like T cell depletion.

Summary

While specific dosing regimens can vary based on the research goal and mouse model, general guidelines suggest:

• In Vivo Depletion: 100 to 500 μg per mouse.
• Flow Cytometry and Other Applications: Tailor dosing based on empirical testing, starting with low concentrations.

Always consult specific literature or product documentation for detailed dosing recommendations tailored to your research needs.