Anti-Mouse/Human CD273 (PD-L2) [Clone 3.2.B8] — Purified in vivo GOLD™ Functional Grade

Clone 3.2.B8 is a monoclonal antibody targeting PD-L2 (CD273), used in in vivo mouse studies to modulate immune responses by blocking the interaction between PD-L2 and PD-1. This blockade plays a role in enhancing T cell activation, proliferation, and cytokine production, potentially leading to increased antitumor immunity.

Key points on in vivo usage:

• Functional Grade: Clone 3.2.B8 is often provided in purified, endotoxin-free preparations suitable for direct injection into mice for experimental manipulation of immune checkpoints.
• Mechanism of Action: By binding to PD-L2 on cells, it prevents the inhibitory signal that PD-L2 delivers through PD-1, effectively releasing the brakes on T cell activity during immune responses.
• Experimental Applications: It is primarily used to study T cell regulation, peripheral tolerance, and mechanisms of immune evasion in tumors. Common in vivo experiments include investigating how blockade of PD-L2 affects:
  • T cell proliferation and activation
  • Cytokine production
  • Tumor growth and immune infiltration

Additional details:

• Species Cross-reactivity: This clone recognizes both mouse and human PD-L2, making it useful in models employing mouse cells and in humanized or xenograft mice.
• Application Examples: Researchers use it in synergy with other checkpoint inhibitors to dissect combinatorial effects on tumor regression and immune cell dynamics. Experimental protocols recommend following product-specific datasheets for dosing and administration.

In summary, clone 3.2.B8 is employed in vivo mainly for immune checkpoint blockade studies in mice, with its primary purpose being to modulate T cell responses by interfering with PD-L2/PD-1 signaling.

The correct storage temperature for sterile packaged Clone 3.2.B8 (anti-mouse/human CD273/PD-L2) is 2-8°C for up to one month if stored sterile as received. For longer-term storage, aliquot aseptically without diluting and store at ? -70°C. Avoid repeated freeze-thaw cycles.

This recommendation is consistent with standard handling for functional grade monoclonal antibodies intended for preclinical use, where short-term storage at refrigerator temperature (2-8°C) is permissible, but deep freezing is preferred for maintaining long-term integrity and activity.

The 3.2.B8 antibody commonly refers to a clone specific for mouse CD273 (PD-L2), used in immunological research, especially for studying immune checkpoints and T cell regulation. In published studies, 3.2.B8 is often used alongside other antibodies or proteins to provide comprehensive immunophenotyping or functional analysis.

Commonly co-used antibodies/proteins with 3.2.B8 (anti-PD-L2) include:

• Anti-CD274 (PD-L1): PD-L1 and PD-L2 are both ligands for PD-1, so studies often examine expression or blockade of both molecules together to assess immunoregulatory pathways.
• Anti-PD-1 (Programmed Death-1): As PD-L2 is a ligand for PD-1, combining 3.2.B8 with anti-PD-1 is standard in immune checkpoint research to analyze the PD-1/PD-L2 pathway.
• Anti-CD80 (B7-1) / CD86 (B7-2): Members of the same B7 family, often analyzed in combination to investigate costimulatory versus coinhibitory signal balance.
• Anti-CD3, Anti-CD4, Anti-CD8: Used to phenotype T cell subsets during functional assays or flow cytometry panels involving PD-L2.
• Anti-MHC class II (e.g., I-A/I-E): As antigen-presenting cells express both MHC-II and PD-L2, co-labeling helps define dendritic cell or macrophage subsets.

These combinations allow researchers to map out immune cell interactions, checkpoint blockade responses, and specific immune cell populations within in vivo or in vitro systems.

In summary, 3.2.B8 (anti-PD-L2) is most frequently used with antibodies targeting PD-1, PD-L1, and standard immunophenotyping markers (CD3, CD4, CD8, CD11c, MHC-II), depending on the precise research application. The exact panel will vary by experiment, but these are the most recurrently reported combinations in immunological literature.

Scientific literature referencing clone 3.2.B8 primarily identifies this monoclonal antibody as a highly specific tool for recognizing PD-L2 (CD273) on both mouse and human cells. The key findings from these citations include:

• Cross-Species Reactivity: Clone 3.2.B8 binds equally well to both mouse and human PD-L2, making it valuable for comparative immunological studies across species.
• Epitope Specificity: It recognizes a distinct epitope on PD-L2, ensuring specific detection and potential functional blockade in assays and in vivo experiments.

Applications in Research:

• Functional Immune Studies: 3.2.B8 is used to explore PD-1/PD-L2 interactions, which play crucial roles in T cell regulation, immune tolerance, and lymphocyte clonal selection.
• Immune Checkpoint Investigation: By interfering with PD-1/PD-L2 binding, clone 3.2.B8 contributes to studies on immune checkpoint blockade, relevant for understanding mechanisms underlying cancer immunotherapy and regulation of T cell responses.

Experimental Utility:

• Versatility in Techniques: The antibody has documented utility in blocking (in vivo), flow cytometry, western blotting, and other biochemical assays, confirming its functional grade and broad applicability in immunological experiments.

No studies from the retrieved results link clone 3.2.B8 directly to infectious disease or TCR clonotype analysis, as discussed in related HIV T cell literature. Instead, its major scientific impact is enabling robust investigations into the function and therapeutic targeting of the PD-1/PD-L2 axis in immune modulation, including autoimmunity and oncology.

If you need details on specific experimental findings or therapeutic outcomes using clone 3.2.B8, additional targeted literature searches may be necessary.