Anti-Mouse CD28 [Clone 37.51] — Purified in vivo GOLD™ Functional Grade

Clone 37.51 is a monoclonal antibody used to target the CD28 costimulatory receptor in mouse studies. It is widely utilized for both in vitro and in vivo applications, particularly for costimulating T cells. Here's how it is used in in vivo mouse studies:

1. CD28 Stimulation: Clone 37.51 can stimulate T cell activation and proliferation by providing a costimulatory signal. This is crucial for enhancing the immune response, as CD28 signaling augments IL-2 production and cytotoxicity of activated T cells.

2. CTL Induction: The antibody helps in the induction of cytotoxic T lymphocytes (CTLs), which are essential for immune responses against pathogens and tumor cells.

3. In Vivo T Cell Activation: By binding to CD28, the antibody can activate T cells in the mouse, mimicking the natural signal provided by CD80 (B7-1) and CD86 (B7-2) ligands.

4. Immune Response Studies: Clone 37.51 is used in studying immune responses, especially in the context of infectious diseases or cancer, where manipulating T cell activation can provide insights into disease mechanisms and potential therapies.

5. Immunotherapy Research: It may be employed in research aimed at enhancing or modulating immune responses for therapeutic purposes, such as in cancer immunotherapy.

When using this antibody in vivo, it is important to select preparations with low endotoxin levels to minimize potential inflammation or immune responses unrelated to the CD28 stimulation.

The correct storage temperature for sterile packaged clone 37.51 (anti-CD28, Syrian Hamster Monoclonal Antibody) is between 2?°C and 8?°C (typically in a standard laboratory refrigerator). Store the antibody solution undiluted and do not freeze.

• Protect from prolonged exposure to light.
• Storage at ambient room temperature is not recommended except during shipping if specified by the supplier.
• Freezing can result in loss of antibody activity and is specifically warned against.
• These guidelines align with standard practices for monoclonal antibody storage.

Always check the product label or accompanying datasheet for any lot-specific recommendations.

The antibody clone 37.51 is used to detect mouse CD28, and in the literature, it is most commonly employed in combination with other antibodies or proteins that identify specific immune cell types, activation markers, and costimulatory molecules.

Typical companions and commonly used antibodies or proteins with 37.51 include:

• CD3: Used to mark T cells, particularly in studies examining T cell activation, as CD3 is a component of the T cell receptor complex involved in signal transduction.
• CD4 and CD8: These antibodies distinguish helper (CD4+) and cytotoxic (CD8+) T cell subsets, providing context for CD28 expression and costimulation effects.
• CD80 (B7-1) and CD86 (B7-2): These are the ligands for CD28 and are often assayed or blocked alongside CD28 to investigate costimulatory pathways in T cell and immune response studies.
• CD25 (IL-2 receptor ?): Frequently used to assess activation status of T cells, as CD28 stimulation via 37.51 augments IL-2 receptor expression.
• Cytokines (e.g., IL-2): Measurement of cytokine production (especially IL-2) is common when analyzing the functional consequences of costimulatory signaling via CD28 engagement.
• Activation markers (CD69, CD44, etc.): Used to further characterize the activation or differentiation state of T cells following costimulation.
• Isotype controls: Such as hamster IgG or hamster IgG-SPRD, used for flow cytometry and other immunoassays to control for nonspecific binding of the 37.51 antibody.

In multicolor flow cytometry panels or in vitro stimulation/activation experiments, antibodies to CD3, CD4, CD8, CD80, CD86, CD25, and cytokine intracellular stains are standard companions to 37.51, enabling detailed immune phenotyping and the study of T cell costimulation mechanisms.

These combinations allow researchers to:

• Dissect T cell subsets and their activation responses
• Analyze costimulatory requirements for CTL induction
• Assess proliferation, cytokine production, and downstream functional effects of CD28 engagement.

Summary Table: Commonly paired antibodies/proteins with 37.51

These combinations are standard in mouse immunology studies examining T cell activation, costimulation, and immune response modulation.

The 37.51 clone is a hamster monoclonal antibody targeting mouse CD28, widely cited in immunological research for its utility in characterizing T cell biology and for functional assays. Key scientific findings from papers citing clone 37.51 include:

• Enhancement of T Cell Proliferation and Cytokine Production: The 37.51 antibody augments in vitro T cell proliferation and cytokine production, demonstrating its role in providing a costimulatory signal necessary for full T cell activation, particularly in synergy with T cell receptor (TCR) engagement.

• Facilitation of Cytotoxic T Lymphocyte (CTL) Development: Studies using 37.51 show it can promote CTL development, underlining the importance of CD28 signaling for immune responses against infected or malignant cells.

• Broad Expression Profile in Mice: The antibody is used for flow cytometric detection of mouse CD28, which is expressed on most thymocytes and at low density on nearly all mature CD4+ and CD8+ T cells, confirming its effectiveness for identifying T cell subsets in mouse models.

• Mechanistic Insights: 37.51 has helped establish that CD28 signaling augments expression of IL-2 and its receptor, increasing T cell proliferation and functionality. This includes its role in boosting T cell cytotoxicity when activated via CD3, as well as providing costimulatory signals essential for optimal immune responses.

• Research Applications: The antibody is applied in:

  • In vitro T cell stimulation and activation
  • In vivo blockage of CD28 to dissect co-stimulation requirements in autoimmune, infectious, and cancer immunology models
  • Detection and phenotyping of CD28 expression on T cells via flow cytometry

• Experimental Evidence: Numerous studies (e.g., Zhang N and He Y-W, J. Exp. Med. 2005; Wang W, J. Immunol. 2007; Barbi J, Blood 2007; Li CR, J Immunol. 2014) rely on 37.51 to dissect costimulatory pathways and the functional consequences for T cell biology, supporting its status as a gold standard for manipulating and identifying CD28 in murine systems.

These findings collectively establish clone 37.51 as a critical tool for dissecting CD28-mediated costimulation in T cell activation, effector differentiation, and immune regulation in murine research.