Anti-Human CD62L – Purified in vivo PLATINUM™ Functional Grade

Clone DREG56 is a monoclonal antibody that specifically targets human CD62L (L-selectin), and in in vivo mouse studies, it is primarily used for functional blocking or depletion of human CD62L, often in models involving human cells or humanized mice rather than native mouse CD62L.

• DREG56 binds to human CD62L, a glycoprotein expressed on human leukocytes (such as neutrophils, monocytes, T, B, and NK cells). CD62L plays a key role in the homing of lymphocytes to peripheral lymph nodes through high endothelial venules (HEVs).
• In vivo uses in mouse studies typically involve humanized mouse models or mice engrafted with human immune cells. DREG56 is used to:
  • Block the function of human CD62L, preventing lymphocyte migration to lymph nodes by inhibiting binding to HEV.
  • Assess the role of CD62L-mediated homing in immune responses, trafficking, or disease models.
• DREG56 does not cross-react with mouse CD62L, so its use in standard mouse models without human cell engraftment is not applicable. For native murine studies, a different anti-mouse CD62L antibody would be required.
• Typical protocols involve systemic administration of DREG56 antibody in mice with human cells, followed by evaluation of immune cell distribution, trafficking, or experimental treatment outcome.

In summary, in vivo mouse use of DREG56 is limited to humanized settings or xenografts, where it is employed to block or study human CD62L-dependent processes. For studies of mouse CD62L, other antibodies are required.

Commonly Used Antibodies and Proteins with DREG56

DREG56 is a monoclonal antibody clone targeting CD62L (L-selectin), widely used in immunology research, especially in studies involving leukocyte adhesion, trafficking, and immune cell phenotyping. While the search results focus on DREG56 itself, several standard antibodies and proteins are commonly used alongside DREG56 in the literature, though not specifically listed in these sources. Based on the broader context of CD62L biology and typical experimental setups, the following are often used with DREG56:

Co-Staining Antibodies

• CD44 antibody: CD44 is functionally linked to CD62L, with overlapping roles in cell adhesion and homing, particularly in lymphocyte migration and disease mechanisms. Co-staining with anti-CD44 helps distinguish cell subsets with different homing potentials (e.g., central vs. effector memory T cells).
• CD4 and CD8 antibodies: These are frequently used in flow cytometry panels to define T cell subsets (helper and cytotoxic) when studying lymphoid cell populations. The cited protocol demonstrates co-staining of CD62L (DREG56) with CD4 using fluorescent conjugates.
• CD19 and CD20 antibodies: Used to identify B cell populations, especially since CD62L is expressed on the majority of B cells. This allows for the delineation of naïve and memory B cell subsets.
• CD14 and CD16 antibodies: For phenotyping monocytes and granulocytes, subsets which also express CD62L.
• Isotype controls: Mouse IgG1 isotype controls, particularly from the same host as DREG56 (mouse IgG1), are essential for setting flow cytometry gates and verifying specificity.
• Secondary detection antibodies: Goat anti-mouse IgG conjugated to fluorophores (e.g., Alexa Fluor® 488) are commonly used for detection when DREG56 is unlabeled.

Functional Assay Proteins

• Glycoprotein ligands (e.g., GlyCAM-1, PSGL-1): Since CD62L is a lectin that mediates adhesion by binding sialylated glycoproteins, these ligands are sometimes used in functional adhesion assays to study CD62L’s interaction with endothelial cells.
• Blocking antibodies: Other selectin family members (e.g., anti-CD62E, anti-CD62P) may be used in co-blocking experiments to dissect the specific contribution of CD62L in leukocyte recruitment.
• Cytokines and chemokines: In vivo and in vitro models often incorporate cytokines (e.g., IL-2, IL-7) or chemokines (e.g., CCL21) to modulate CD62L expression and function on lymphocytes.

Negative Controls

• Murine cells: When characterizing human samples, murine cells can serve as negative controls, provided there is no cross-reactivity between species.
• Isotype controls: As noted, these are critical for ensuring signal specificity in both flow cytometry and functional assays.

Summary Table

These combinations are standard in immunology research focused on lymphocyte homing, adhesion, and migration, reflecting the central role of CD62L (detected by DREG56) in these processes. Always check for cross-reactivity and optimize panels based on specific experimental goals.

Based on the scientific literature cited across multiple research applications, clone DREG56 has generated several key findings that have advanced our understanding of CD62L/L-selectin biology and immune function.

Cellular Distribution and Expression Patterns

The DREG56 clone has been instrumental in characterizing the expression profile of CD62L across immune cell populations. Research has established that CD62L is expressed on neutrophils, monocytes, T- and B-lymphocyte subsets, and NK cells. The antibody recognizes a 74-95 kDa type I transmembrane glycoprotein that serves as a critical cell adhesion molecule on most circulating leukocytes.

Lymphocyte Homing and Tissue Selectivity

One of the most significant discoveries using DREG56 is its role in defining lymphocyte homing mechanisms. The clone specifically inhibits over 90% of binding between human lymphocytes and high endothelial venules (HEV) in frozen sections of peripheral lymphoid tissue, but notably not in mucosal lymphoid tissue. This finding established L-selectin as a human lymphocyte homing receptor with tissue-specific functions, particularly for peripheral lymph node HEV targeting.

Clinical Applications in Disease Research

The literature reveals extensive use of DREG56 in clinical research across multiple disease contexts:

Cancer Research: Studies have utilized the clone to investigate immune profiles in cancer patients, with research examining cellular immune responses and potential therapeutic targets.

Pediatric Medicine: Research has employed DREG56 to study immune development and function in pediatric populations, contributing to understanding age-related immune system variations.

Infectious Disease Studies: The clone has been valuable in examining immune responses during infections, including detailed flow cytometric analysis of leukocyte populations during disease progression.

COVID-19 Research Applications

Recent pandemic research has leveraged DREG56 for comprehensive immune profiling in COVID-19 patients. Studies have used high-dimensional flow cytometry with this clone to examine alterations in innate and adaptive immune compartments, revealing key myeloid signatures associated with severe disease. This work has contributed to understanding how immune cell characteristics and relative abundance change during COVID-19 progression.

Technical Validation Across Multiple Applications

The scientific literature demonstrates DREG56's versatility across various experimental techniques. Citations show successful applications in flow cytometry, western blotting, immunohistochemistry on frozen sections, and immunofluorescence. The clone has been validated for both research and clinical applications, with studies consistently demonstrating its reliability for detecting CD62L expression changes across different experimental conditions.

These findings collectively establish DREG56 as a critical research tool that has significantly advanced our understanding of immune cell trafficking, homing mechanisms, and disease-associated immune alterations across diverse clinical and research contexts.

There is no standardized in vivo dosing regimen for clone DREG56 (anti-human CD62L/L-selectin) across mouse models documented in the provided results. The available sources describe DREG56 primarily as a research-use-only (RUO) reagent for flow cytometry to detect human CD62L expression on human peripheral blood lymphocytes, not as a therapeutic or depleting antibody administered to living mice in preclinical models.

Key Context and Clarifications:

• Dosing Regimens Not Reported: None of the retrieved sources specify intraperitoneal, intravenous, or other in vivo dosing instructions for clone DREG56 in mouse models, in contrast to well-characterized mouse-specific or functional antibodies such as anti-PD-1 or anti-CD3.

• Usage in Research: DREG56 is routinely cited in protocols for ex vivo or in vitro applications—especially for flow cytometric staining of human cells, including those extracted from humanized mice. The protocols typically use very low amounts (e.g., 5–20 ?l of antibody per test for ~10? cells) and are not intended as dosing regimens for systemic administration.

• Mouse Models and DREG56: In studies utilizing humanized mice (such as NSG mice engrafted with human PBMCs), DREG56 is used to analyze the expression of human CD62L on human T cells in mouse tissues by flow cytometry, not for in vivo modulation or depletion of target cells. No dosing regimens analogous to those for antibodies like anti-PD-1 (e.g., 100–200 ?g/mouse per injection) are described for DREG56.

• Other Antibody Dosing: For reference, antibodies used functionally in mice (e.g., for depletion or signaling modulation) have published dose ranges and schedules. For example, anti-PD-1 antibodies are typically used at 100–500 ?g per mouse, often given intraperitoneally every 3–4 days, tailored for mouse immune targets and not for human-specific targets such as CD62L.

Summary Table: DREG56 vs. Common Functional Antibodies in Mouse Models

If you seek in vivo dosing regimens for DREG56 in humanized mouse models or other contexts, this is not supported by the current literature or datasheets; DREG56 is not used for in vivo functional intervention in mice. Any proposed systemic use would require novel validation and justification.

If you meant flow cytometry staining protocols or require guidance for other anti-CD62L clones used in mouse (cross-reactive) models, please clarify.