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

Clone DREG56 is a mouse monoclonal antibody that recognizes human CD62L (L-selectin), commonly used to study human immune cell populations. In vivo applications of DREG56 in mice occur almost exclusively in humanized mouse models—mice engrafted with human immune cells (e.g., PBMCs or hematopoietic stem cells)—rather than conventional mice, as the antibody is specific for the human protein and does not cross-react with mouse CD62L.

Common in vivo applications of DREG56 in mice:

• Tracking or detecting human lymphocyte homing: DREG56 blocks and detects human CD62L, which mediates lymphocyte rolling on high endothelial venules (HEV) and thus cell homing to lymphoid tissues. It has been used to inhibit human lymphocyte adhesion and migration in humanized mice.
• Flow cytometry analysis: DREG56 is routinely used to quantify and phenotype human CD62L+ leukocytes in the blood and tissues of humanized mice, serving as a marker for naïve and central memory T cells.
• In vivo inhibition/blockade studies: Functional (low endotoxin, azide-free) DREG56 can be used for blocking CD62L-mediated interactions, assessing the role of human CD62L in cellular trafficking, inflammation, or disease models within humanized mice.
• Evaluation of immune reconstitution: In humanized mice, DREG56 helps assess the presence, frequency, and phenotype of engrafted human leukocytes, including tracking T, B, and NK cell populations.

Supporting details:

• DREG56 does NOT react with native mouse CD62L; use in conventional mouse models will not yield specific data unless human CD62L-expressing cells (e.g., from human PBMCs or engineered stem cells) are present.
• Blocking activity has been reported for use in both in vitro and in vivo settings, where DREG56 is used to dissect the role of human L-selectin in cell trafficking and immune responses in humanized mouse models.

If your aim is to manipulate or study endogenous mouse L-selectin, a mouse-reactive antibody clone (like MEL-14) is required, as DREG56 is not cross-reactive.

Summary Table: DREG56 in Humanized Mouse Applications

In conclusion, clone DREG56 is utilized in vivo in mice almost exclusively for research involving engrafted human immune cells, particularly for tracking, blocking, or phenotyping human CD62L+ leukocytes. It does not serve in murine-only in vivo biology.

When used in conjunction with the DREG56 antibody for CD62L, researchers often employ other antibodies or proteins to analyze various immune cell populations and their interactions. Here are some commonly used antibodies or proteins:

1. CD3 Antibodies: These are used to identify T cells, which are crucial for immune responses. CD62L is expressed on naive T cells, so combining CD3 with DREG56 can help differentiate between naive and memory T cells.

2. CD4 and CD8 Antibodies: These are used to distinguish between helper T cells (CD4+) and cytotoxic T cells (CD8+), allowing for more detailed analysis of T cell subsets.

3. CD19 Antibodies: These identify B cells, which also express CD62L. This combination helps in studying B cell subsets and their interactions.

4. CD11b Antibodies: These are used to identify monocytes and macrophages, which express CD62L. This combination is valuable for studying myeloid cell populations.

5. CD34 Antibodies: CD34 is a ligand for CD62L and is involved in hematopoietic stem cell differentiation. Using CD34 with DREG56 can help in understanding hematopoiesis and stem cell homing.

6. ICAM-1 and VCAM-1 Antibodies: These molecules are involved in later stages of leukocyte adhesion and are often studied in conjunction with adhesion molecules like CD62L to understand the full spectrum of leukocyte recruitment.

These combinations allow researchers to dissect the complex interactions between different immune cell types and their roles in inflammation, homing, and hematopoiesis.

Clone DREG56 (also known as DREG-56) is a monoclonal antibody used to detect human CD62L (L-selectin), a cell adhesion molecule primarily expressed on neutrophils, monocytes, and subsets of T and B lymphocytes. The key findings in the scientific literature and usage of this clone relate to its specificity, its applications in immunophenotyping and its pivotal role in studies of immune cell subsets and migration:

• Specific Target: DREG56 is specific for CD62L/L-selectin, a 65–76 kDa single-chain glycoprotein crucial for leukocyte homing to lymphoid tissue and recruitment to sites of inflammation.

• Cell Populations Identified: It is used to identify CD62L on neutrophils, monocytes, B cells, and notably naïve and central memory T cells. In immunology research, DREG56 allows distinction between CD62L^high and CD62L^low effector/memory T cell subsets.

• Applications: DREG56 is widely used in:

  • Flow cytometry and immunofluorescence to profile leukocyte populations.
  • Sorting and isolation of lymphocyte populations based on homing and activation status.
  • Studying cell migration, immune surveillance, and lymphocyte trafficking.

• Phenotyping Immune Responses: The antibody has contributed to research on:

  • The functional heterogeneity of T and NK cell populations, such as investigating shifts in CD62L expression in adaptive immune responses and after infections like CMV.
  • Identification of specific myeloid and lymphoid signatures associated with disease states and immune activation.

• Technical Note: DREG56 detects an extracellular epitope of CD62L and is verified not to cross-react with mouse L-selectin, making it reliable for human studies.

• Vendor and RRID information: DREG56 is a standard tool antibody available from multiple vendors, including Thermo Fisher, Bio-Techne, and BioLegend, referenced by RRID:AB_3699478 and others for reproducibility.

Literature does not report off-target effects or loss of specificity, and there is strong consensus that DREG56 is a gold-standard reagent for human CD62L identification.

In summary: Clone DREG56 is foundational in immune cell phenotyping—especially T cell subset classification, immune monitoring in disease and vaccination, and research on cell migration and immune surveillance—due to its high specificity and consistent performance in detecting human CD62L/L-selectin.

Dosing regimens of clone DREG56 (an anti-human CD62L/L-selectin antibody) are primarily established for ex vivo and in vivo detection or depletion of human CD62L, and the optimal dose can vary based on the mouse model, application, and readout sensitivity.

Key points on dosing regimens:

• Flow cytometry in humanized mouse models: For in vivo or ex vivo flow cytometric analysis of human cells (e.g., PBMCs) engrafted in NSG or other immunodeficient mice, typical recommendations are ≤0.125 µg per million cells in 100 µL staining volume.
• Application context: DREG56 is generally used for detection rather than functional depletion—thus, in most published mouse models, it's applied to isolated cells for flow cytometry after tissue harvest.
• Species restriction: DREG56 does not bind mouse CD62L; thus, in non-humanized mouse models, it is generally not employed except as a negative control or in cross-species studies.
• Humanized models: In NSG or similar humanized mice, dosing follows guidance for antibody staining (e.g., ≤0.125 µg/million cells), not systemic injection for modulation or depletion. Engraftment-dose optimization pertains to PBMCs (e.g., 8–10 × 10⁶ PBMCs) rather than DREG56 antibody.
• Functional studies: For in vivo perturbation or depletion, there is little evidence or protocol for systemic administration of DREG56 in mice, as the antibody is not generally reported to be used for functional in vivo blockade but rather for phenotyping.

Table: DREG56 usage in common mouse model settings

Additional context:

• Suppliers (BioLegend, BD, ThermoFisher, eBioscience) recommend optimizing the antibody concentration for individual experimental setups.
• There is no standard published in vivo dosing regimen for DREG56 equivalent to depleting/dosing antibodies used for checkpoint blockade or cell depletion (e.g., anti-CD3, anti-Ly6G) in mice.

Summary:
DREG56 dosing is not varied for systemic dosing across different mouse models; it is mainly used in ex vivo stainings for human cell detection in humanized mice, typically at ≤0.125 µg/million cells per 100 µL, with species specificity restricting utility outside of humanized models.