Anti-Mouse LPAM-1 – Purified in vivo GOLD™ Functional Grade

Applications of DATK32 Clone in In Vivo Mouse Studies

Clone DATK32 refers to a monoclonal antibody specifically targeting mouse LPAM-1, also known as integrin ?4?7. This integrin is expressed on peripheral lymphocytes, certain thymocytes, and bone marrow progenitors, and is crucial for lymphocyte adhesion and migration to gut-associated lymphoid tissues via binding to ligands like VCAM-1, MAdCAM-1, and fibronectin.

In Vivo Functional Use

• Blockade of Integrin ?4?7 (LPAM-1) Function: The DATK32 antibody is widely used in vivo to neutralize the function of integrin ?4?7, effectively blocking lymphocyte adhesion and migration mediated by this receptor. This is particularly relevant in studies of intestinal immunity, inflammation, and lymphocyte trafficking.
• Prevention of Lymphocyte Homing: By administering DATK32 in vivo, researchers can investigate the role of ?4?7 in diseases such as inflammatory bowel disease (IBD) or intestinal infections, where lymphocyte recruitment to the gut is a key pathological feature. The antibody’s ability to block adhesion to MAdCAM-1 (the primary gut-homing ligand) underlies its effectiveness in these models.
• Neutralization Studies: DATK32 is validated for in vivo neutralization, meaning it can be injected into live mice to observe the physiological effects of blocking ?4?7, including changes in immune cell distribution, inflammation, and disease outcomes.

Experimental Considerations

• Isotype and Controls: DATK32 is a rat IgG2a, ? antibody. Appropriate isotype controls (e.g., InVivoMAb rat IgG2a isotype control) should be used to distinguish specific effects from nonspecific antibody responses.
• Administration: The antibody is typically administered via intraperitoneal or intravenous injection. The exact dose and regimen depend on the experimental model and should be optimized for each study.
• No Stabilizers/Preservatives: The antibody is supplied in PBS without stabilizers or preservatives, and should not be frozen but stored at 4°C.
• Purity and Safety: The product is >95% pure, with low endotoxin levels, making it suitable for in vivo use.

Supported by Peer-Reviewed Literature

DATK32’s efficacy in blocking ?4?7-mediated adhesion in vivo is supported by multiple peer-reviewed studies, confirming its utility for investigating lymphocyte trafficking and gut immunity in mouse models. References to these studies are typically provided by suppliers (e.g., BioLegend) for further methodological detail.

Summary Table: Key Features of DATK32 in In Vivo Studies

Conclusion

Clone DATK32 is a critical tool for in vivo mouse studies focused on dissecting the role of integrin ?4?7 in immune cell trafficking, particularly to the gut. Its ability to specifically and effectively block this integrin in vivo enables researchers to investigate mechanisms of intestinal inflammation, immunity, and related pathologies.

The correct storage temperature for sterile packaged clone DATK32 depends on the intended storage duration and specific formulation.

Short-term Storage

For short-term storage of up to one month, clone DATK32 should be stored at 2-8°C. This applies to functional grade preclinical antibodies that are aseptically packaged and formulated.

Long-term Storage

For long-term storage, the antibody should be stored at ? -70°C. When storing for extended periods, it's recommended to aseptically aliquot the antibody into working volumes without diluting to avoid repeated freeze-thaw cycles.

Additional Storage Considerations

Some formulations specify storage at -10°C to -25°C, while others recommend storing at 2-8°C undiluted and protected from light. The antibody should always be stored in the dark and should not be frozen for certain formulations.

Important handling recommendations include centrifuging the vial and gently mixing the solution upon receipt before removing the cap, and avoiding repeated freeze-thaw cycles for optimal antibody stability.

Antibodies Commonly Used Alongside DATK32

The DATK32 antibody is a monoclonal antibody specific for the mouse integrin ?4?7 heterodimer (LPAM-1), primarily used to block or detect the function and expression of this integrin in immune cell adhesion and migration studies. In the literature, DATK32 is frequently used in combination with other antibodies and proteins to study leukocyte trafficking, particularly in mucosal immunology and inflammation models.

Core Ligands and Adhesion Molecules

DATK32’s primary biological targets are the ligands for integrin ?4?7, which are central to its functional studies:

• VCAM-1 (Vascular Cell Adhesion Molecule-1): A major endothelial ligand for ?4?7, involved in lymphocyte recruitment to inflamed tissues.
• MAdCAM-1 (Mucosal Addressin Cell Adhesion Molecule-1): Expressed on gut endothelium, essential for lymphocyte homing to intestinal mucosa; binding to ?4?7 is a key step in gut-specific immune responses.
• Fibronectin: An extracellular matrix protein that also serves as a ligand for ?4?7, contributing to cell adhesion and migration.

Co-Staining and Flow Cytometry

In flow cytometry (FACS) experiments, DATK32 is often used alongside antibodies against other cell surface markers to identify specific leukocyte subsets or activation states. For example:

• CD45R (B220): A pan-B cell marker used to identify B lymphocytes in co-staining experiments, as demonstrated in bone marrow cell analyses.
• Other lineage markers: While not explicitly listed in the provided sources, it is common in the literature to co-stain with CD3 (T cells), CD4, CD8, CD19, or CD11b to distinguish lymphocyte subsets or myeloid cells in complex tissues.

Functional Blocking and Neutralization Studies

DATK32 is used in functional assays to block ?4?7-mediated adhesion. In these contexts, the following are relevant:

• Isotype controls: Rat IgG2a isotype controls are recommended to confirm specific staining or blocking by DATK32.
• Integrin-neutralizing antibodies: While DATK32 itself is the primary blocker for ?4?7, studies may include antibodies against other integrins (e.g., anti-?4, anti-?7) or their ligands to dissect specific adhesion pathways.

In Vivo Models and Therapeutic Context

In preclinical studies, especially those modeling inflammatory bowel disease (IBD), DATK32 is sometimes used in parallel with:

• Humanized anti-?4?7 antibodies (e.g., vedolizumab): These are used to compare mechanisms across species or to benchmark the efficacy of new therapeutics.
• Cytokines and chemokines: To study the role of ?4?7 in lymphocyte recruitment, experiments may include stimulation with TNF-?, IL-12, or CCL25 (TECK), which are implicated in gut homing.

Summary Table

References to Key Studies

Several foundational papers are cited in the context of DATK32 usage, including studies on lymphocyte adhesion blocking (Andrew et al., Berlin et al., Berlin-Rufenach et al.), immunoprecipitation (Rivera-Nieves et al.), and immunohistochemistry (Bogetto et al.). These studies typically employ DATK32 alongside the ligands and markers listed above to dissect the role of ?4?7 in immune cell trafficking.

Conclusion

DATK32 is most commonly used alongside its natural ligands (VCAM-1, MAdCAM-1, fibronectin), lineage-specific antibodies (e.g., CD45R), isotype controls, and sometimes therapeutic analogs (e.g., vedolizumab) in both in vitro and in vivo studies of lymphocyte adhesion and mucosal immunity. The choice of co-reagents depends on the experimental design, but the above list covers the most prevalent combinations in the literature.

Clone DATK32 is a monoclonal antibody specific for mouse integrin ?4?7 (LPAM-1), widely used to block ?4?7-mediated lymphocyte adhesion and trafficking. Scientific literature highlights several key findings from studies that cite and use clone DATK32:

Key Findings:

• Effective Blocking of Integrin ?4?7 Functions:

  • DATK32 blocks ?4?7-mediated adhesion of lymphocytes to VCAM-1, MAdCAM-1, and fibronectin both in vitro and in vivo. This property is fundamental for studies manipulating lymphocyte migration, especially in models of mucosal immunology.

• Targeted Delivery Improves Local Tissue Exposure:

  • Intracolic (IC) administration of DATK32 leads to much higher concentrations of the antibody in colon tissues and contents than intraperitoneal (IP) administration, while keeping systemic exposure low. This translates to:
    • Prolonged tissue-level exposure (elevated for up to 24h) post-dosing
    • Potential for improved therapeutic responses in localized inflammation models, such as IBD.

• Reduction in Pathogenic Lymphocyte Populations:

  • DATK32 significantly reduces ?4?7-expressing memory T-cells in the Peyer’s patches and mesenteric lymph nodes following targeted delivery in mouse models of acute colitis. This demonstrates its immunomodulatory potential by directly affecting gut-homing T-cell populations.

• Topical Antibody Delivery Suggests Improved IBD Therapy:

  • Studies have shown that localized (topical) DATK32 treatment results in greater pharmacodynamic effects and greater reductions in inflammation compared to systemic dosing, supporting the idea of targeted delivery for better clinical outcomes in inflammatory bowel disease.

• Cell Adhesion and Migration Research:

  • DATK32 is recognized for its unique specificity in cell adhesion and migration assays, allowing researchers to precisely interrogate ?4?7’s role in immune cell trafficking. The epitope recognized by DATK32 is conformational, facilitating robust detection and manipulation of integrin function.

Supporting Context and Additional Notes:

• Immunohistochemistry/Immunoprecipitation: DATK32 is used for both immunoprecipitation and immunohistochemical staining of frozen tissue sections to localize and assess ?4?7 distribution in tissues.
• Antigen and Isotype: DATK32’s antigen targets include LPAM-1, with alternative names such as CD49D, integrin alpha 4 subunit, and ITGA4.
• Host/Isotype: The antibody is a rat IgG2a.

Summary Table: Main Functional Applications of DATK32

These results collectively establish clone DATK32 as the benchmark for selective blocking and detection of ?4?7 integrin in immunological research, facilitating studies of lymphocyte trafficking, mucosal immunity, and targeted antibody therapy for conditions such as IBD.