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

The clone 30-H12 is used in in vivo mouse studies primarily to deplete T lymphocytes in mice expressing the Thy1.2 (CD90.2) antigen. This monoclonal antibody binds specifically to mouse Thy1.2, a surface glycoprotein found on thymocytes, mature T cells, certain hematopoietic stem cells, neurons, epithelial cells, and fibroblasts of Thy1.2-expressing mouse strains (e.g., C57BL/6, BALB/c).

Key in vivo applications of clone 30-H12 include:

• Targeted depletion of T cells: By binding to Thy1.2, the antibody can be used to deplete T cells in experimental mouse models, which is useful for studying immune cell function, transplantation tolerance, and autoimmune diseases.
• Functional modulation: The antibody has been shown to induce calcium flux in thymocytes and, when used with anti-CD3/TCR antibodies, can promote thymocyte activation and apoptosis.
• Cell tracking and identification: In flow cytometry and immunohistochemistry, 30-H12 is also commonly used to detect or isolate Thy1.2-positive cells from tissues, often as part of studies involving hematopoietic stem cells or neuronal cells.
• Specificity to Thy1.2 strains: The antibody does not cross-react with mice expressing Thy1.1 (such as AKR/J, PL strains), so its depletion/application is limited to Thy1.2-expressing strains.

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

In experimental design, 30-H12 is administered in vivo (often IV or IP) to deplete T cells or label Thy1.2+ populations, and its use has been validated for functional depletion and identification by several commercial and research sources.

If you need details on administration protocols or specific model examples, please specify the intended application (e.g., depletion vs. tracking).

Commonly used antibodies and proteins with 30-H12 (anti-mouse CD90.2/Thy1.2) in the literature include:

• CD45: CD90.2 is known to interact with CD45, and antibodies against CD45 are frequently used alongside 30-H12, especially in combination experiments to study signal transduction and T cell biology.
• CD3/TCR complex (CD3, TCR?, etc.): 30-H12 is often paired with antibodies targeting components of the T cell receptor (TCR) and CD3, for costimulation studies and characterization of T cell populations.
• CD16/CD32 (Fc Block, clone 93): Pre-incubation with anti-mouse CD16/CD32 is recommended to block Fc receptors and reduce non-specific binding during flow cytometry or immunofluorescence staining.
• Other T cell subset markers: 30-H12 is commonly used alongside antibodies to CD4 and CD8 for phenotyping subpopulations among thymocytes and peripheral T cells.
• Isotype control antibodies: Rat IgG2b isotype controls are used as negative controls in experiments with 30-H12.
• Stem cell and lineage markers: Because CD90.2 is also a marker for hematopoietic stem cells and some neuron populations, researchers may co-stain with lineage markers such as Sca-1, c-Kit, and neural cell markers depending on the biological context.

Relevant applications for these combinations include:

• Flow cytometry
• Immunohistochemistry (IHC) and immunofluorescence
• T cell depletion and subset isolation
• Costimulation and activation assays

In multiparameter flow cytometry, panels with 30-H12 frequently include antibodies against CD3, CD4, CD8, CD45, CD44, CD62L, and CD25. The exact combinations vary with the experimental design.

Blocking reagents (anti-CD16/CD32) and isotype controls are essential for accurate gating and data interpretation when using 30-H12 in multicolor panels.

Clone 30-H12 is a well-characterized monoclonal antibody that specifically binds to CD90.2 (Thy-1.2), a cell surface glycoprotein found on mouse T cells and several other cell types, and is frequently used in immunology research to label, deplete, or functionally manipulate Thy-1.2+ cells in mouse models.

Key findings and uses reported in the scientific literature include:

• Specificity: 30-H12 binds specifically to the Thy-1.2 alloantigen (CD90.2) on most mouse strains, distinguishing Thy-1.2+ from Thy-1.1+ cells (e.g., not cross-reacting with AKR/J or PL mouse Thy-1.1, or rat Thy-1). It is expressed on thymocytes, most T lymphocytes, some intraepithelial T lymphocytes, some epithelial cells, fibroblasts, neurons, and hematopoietic stem cells, but not B lymphocytes.

• Functional studies: The antibody is commonly used to selectively deplete or identify Thy-1.2+ populations. For instance, depletion using 30-H12 was crucial in studies showing that protection against Mycobacterium tuberculosis vaccination in CD4-deficient mice depends on Thy-1.2+ double-negative (CD4– CD8–) T cells rather than conventional T cell subsets. Mice treated with 30-H12 lost vaccine-induced protection in both lungs and spleens, confirming the effector role of Thy-1.2+ T cells in this context.

• Mechanistic insights: Crosslinking the 30-H12 antibody on thymocytes can induce calcium influx, and co-crosslinking with anti-CD3 enhances signal transduction, promotes apoptosis of thymocytes, and inhibits the CD3-mediated proliferative response of mature T lymphocytes. This has made 30-H12 an important tool for dissecting T cell signaling and homeostasis.

• Technical applications: 30-H12 is widely validated for flow cytometry, cell preparation, and in vivo depletion. Researchers often use the antibody to verify T cell subsets, purify populations, or perform depletion experiments. The antibody's performance in these settings is supported by multiple suppliers and literature reports.

• General relevance: The discoveries made possible with 30-H12 (especially regarding double-negative T cells and T cell signaling) have broad implications for understanding immune responses, vaccine design in immunocompromised hosts, and mouse immunophenotyping.

In summary, 30-H12 is a critical reagent for mouse T cell studies, enabling precise identification and manipulation of Thy-1.2+ cells, elucidating T cell subset functions, and mediating important discoveries in immunology.

Dosing regimens of clone 30-H12 (anti-mouse CD90.2/Thy1.2) vary depending on the application, mouse strain, and specific experimental design, but typical practices are as follows:

• Common Uses: 30-H12 is primarily used in vivo for depletion of T lymphocytes in mouse models expressing the Thy1.2 (CD90.2) alloantigen (e.g., strains like C57BL/6, BALB/c) and for functional studies such as costimulation and apoptosis assays.

• General Dosing Guidance: While few sources specify exact regimens in open product listings, typical depletion protocols for similar antibodies use doses of 100–500 ?g per mouse, frequently delivered via intraperitoneal injection. Multiple doses are common, administered every 3–4 days, especially when sustained depletion is required.

• Strain Specificity: The efficacy of 30-H12 depends on the genetic background:

  • Responsive strains (Thy1.2 positive): Includes C57BL/6, BALB/c, CBA, C3H, SJL, DBA; for these, depletion is effective using 30-H12.
  • Non-responsive strains (Thy1.1 positive): Such as AKR/J, PL; 30-H12 does not cross-react with these and thus is ineffective.

• Reported Applications:

  • Depletion: Used for robust T-cell depletion in responsive strains (methodology aligns with standard in vivo antibody protocols).
  • Functional Assays: For costimulation, cytometry, and apoptosis, lower doses compatible with analytical detection rather than depletion are typical.
  • Route: Intraperitoneal injection is standard for in vivo depletion, though intravenous injection may be used in some protocols.

• Product Concentrations and Storage: Antibody is typically supplied at concentrations like 0.1 mg/mL, to be stored at 2–8°C and protected from light; dosing volumes are calculated accordingly to achieve the desired mass per animal.

• Experimental Customization: Actual dosing schedules, frequency, and total amount may be adjusted based on:

  • The mouse model’s size, age, and experimental aim
  • Desired duration and completeness of T-cell depletion
  • Combination with other antibodies or treatments
  • Published protocols in the relevant literature for the exact use case

Summary Table: Typical Dosing Parameters for Clone 30-H12

Note: For precise dosing regimens matched to specific mouse models and your experimental needs, consult relevant primary literature using clone 30-H12 or contact product technical support at suppliers like Bio X Cell or Leinco, as peer-reviewed references often provide optimized schedules for various disease or depletion models.

No direct citations in the results provided a specific experimental dosing schedule for clone 30-H12 across multiple mouse models, but guidelines above reflect typical antibody depletion practices and known specificity of the clone.