Anti-Mouse CD48 (Clone HM48-1) – Purified in vivo GOLD™ Functional Grade

Use of Clone HM48-1 in In Vivo Mouse Studies

The HM48-1 monoclonal antibody specifically binds to mouse CD48, a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that is broadly expressed on leukocytes and plays roles in immune cell adhesion and activation. CD48 interacts mainly with CD2 and CD244 (2B4), molecules involved in T cell and natural killer (NK) cell activation.

Functional Effects of HM48-1 in Vivo

• Blocking CD48 Interactions: HM48-1 is widely used to block the binding of CD48 to its ligands CD2 and CD244 in vivo, thereby inhibiting downstream signaling pathways important for immune cell communication and activation.
• Modulation of Immune Responses: Administration of HM48-1 has been shown to inhibit the proliferative response of mitogen-activated spleen cells and can augment the proliferative response of T cells when cross-linked with anti-CD3 antibodies. It also inhibits cytotoxic T lymphocyte (CTL) priming in vitro, which is relevant for understanding immune regulation in vivo.
• Transplantation Studies: HM48-1 is reported to prolong the survival of cardiac allografts in mice, an effect that is significantly enhanced when combined with anti-CD2 antibodies. This suggests a role for CD48 in transplant rejection and demonstrates the utility of HM48-1 as a tool in transplantation immunology studies.

Typical Experimental Applications

• Immune Modulation: To study the role of CD48 in adaptive immune responses, HM48-1 can be administered to mice to block CD48-mediated interactions and observe effects on T cell, B cell, and NK cell activities.
• Transplantation Models: In cardiac allograft models, HM48-1 is used to determine the contribution of CD48 to graft rejection and to explore potential therapeutic strategies for prolonging graft survival.
• Infection and Inflammation: While not explicitly detailed in the provided results, blocking CD48 in vivo could also be relevant in models of infection or autoimmune disease, given CD48's central role in leukocyte adhesion and signaling.

Practical Considerations

• Antibody Purity: For in vivo studies, it is recommended to use low-endotoxin, purified forms of HM48-1 (such as Ultra-LEAF™ grade) to avoid confounding effects from endotoxin contamination.
• Dosing and Administration: Specific protocols for in vivo use (dose, route, frequency) should be determined empirically and may vary depending on the experimental model. Manufacturer instructions and published literature should be consulted for optimal results.
• Specificity: HM48-1 is specific to mouse CD48 and does not cross-react with rat leukocytes, ensuring targeted effects in mouse models.

Summary Table: In Vivo Applications of HM48-1 in Mice

Conclusion

HM48-1 is a valuable tool for in vivo mouse studies investigating the role of CD48 in immune regulation, transplantation, and potentially other immune-mediated processes. Its primary use is to block CD48 interactions with CD2 and CD244, thereby modulating immune cell function and influencing outcomes in experimental models of immunity and transplantation.

The correct storage temperature for the sterile packaged clone HM48-1 is between 2°C and 8°C (36°F and 46°F). It should be stored undiluted and protected from prolonged exposure to light. Do not freeze the antibody solution. However, some sources suggest a more specific storage at 4°C for certain formulations. Always check the specific product documentation for precise storage instructions.

Other commonly used antibodies or proteins in the literature with HM48-1 (anti-mouse CD48) include those targeting the CD2 and CD244 (2B4) receptors, as these are principal ligands and interacting partners of CD48 on lymphocytes. Researchers frequently combine HM48-1 with antibodies against surface markers such as CD3, CD4, CD8, CD19, and CD45 to identify specific T cell, B cell, and hematopoietic populations in flow cytometry panels or functional assays.

Key antibodies and proteins commonly co-used with HM48-1:

• Anti-CD2: As CD2 is a known ligand for CD48, blocking or detecting CD2 is common in studies focusing on cellular interactions and signaling.
• Anti-CD244 (2B4): CD244 is another receptor for CD48 and is particularly relevant in investigations of NK cell and T cell regulation.
• Lineage markers: These include anti-CD3 (T cells), anti-CD4, anti-CD8 (T cell subsets), anti-CD19 (B cells), and anti-CD45 (pan-leukocyte) antibodies for immunophenotyping and functional studies.
• Activation/functional markers: Antibodies against proteins like CD69, CD25, or cytokines (e.g., IFN?, TNF?) may also be co-stained to study cell activation in the context of CD48-mediated signaling.

These combinations are essential for multiparametric flow cytometry, cell sorting, and mechanistic studies of immune cell function and interaction, especially in mouse models of immunology and inflammation.

Based on the available information, clone HM48-1 is a widely used research antibody with significant scientific applications, though specific key findings from individual citations are limited in the provided sources.

Clone HM48-1 Characteristics and Applications

Clone HM48-1 is an Armenian hamster anti-mouse CD48 monoclonal antibody that recognizes an epitope on mouse CD48. CD48 is also known by several alternate names including BCM1, B-cell Membrane Protein, and BLAST1. The antibody has an IgG isotype and interacts with CD2 and CD244 as ligand/receptors.

Research Usage and Citation Frequency

The antibody has demonstrated substantial research impact, with 35 publications citing its use in scientific studies. This citation frequency indicates its importance as a research tool in immunological studies. The antibody is commonly used in multiple experimental applications including flow cytometry (FC), blocking (B), immunoprecipitation (IP), costimulation assays, functional assays (FA), and in vivo studies.

Technical Applications in Research

For flow cytometry applications, the recommended concentration is ?1.0 ?g per 10? cells in a volume of 100 ?l or 100 ?l of whole blood. The antibody is available in various formats including FITC conjugates for fluorescence applications and purified functional grade formulations for in vivo studies.

Quality and Purity Standards

The functional grade version maintains high purity standards with ?95% monomer by analytical size exclusion chromatography and >95% by SDS-PAGE. Endotoxin levels are kept extremely low at ?1.0 EU/mg, making it suitable for in vivo functional studies.

The antibody is manufactured using animal-free in vitro cell culture techniques and purified through multi-step processes including Protein A or G chromatography to ensure low levels of contaminants and aggregates. This high-quality production makes HM48-1 a reliable tool for immunological research applications.