Anti-Mouse CD62L [Clone MEL-14] — Purified in vivo PLATINUM™ Functional Grade

Clone MEL-14, a rat monoclonal antibody targeting mouse CD62L (L-selectin), is widely used in various in vivo applications involving mice. These applications include:

1. Functional Blockade of Lymphocyte Homing: MEL-14 is used to block lymphocyte homing to lymph nodes. It prevents the migration of lymphocytes by inhibiting their binding to high endothelial venules (HEV) in peripheral lymph nodes.

2. Immunological Research: It is employed in studies aimed at understanding immune cell migration, particularly in the context of inflammatory processes and lymphocyte trafficking between blood and lymphoid tissues.

3. Flow Cytometry and Immunohistochemistry: MEL-14 is used as a surface marker for identifying lymphocyte subpopulations in flow cytometry and for immunohistochemical staining to study tissue distribution of cells expressing CD62L.

4. In Vivo Depletion and Blocking Experiments: This clone is utilized in experiments to deplete or block specific cell populations, such as neutrophils or subsets of T cells, allowing researchers to study the role of these cells in immune responses.

Overall, MEL-14 is a versatile tool for studying lymphocyte homing, immune cell migration, and the role of selectins in immune responses within mouse models.

MEL-14, a rat monoclonal antibody against mouse CD62L (L-selectin), is frequently used alongside several other antibodies and proteins in immunological research. These companion markers help researchers comprehensively characterize immune cell populations and their functional states.

T and B Cell Markers

CD3 serves as a pan-T cell marker and is commonly used with MEL-14 to identify and analyze T cell populations. Since CD62L expression varies between naïve and memory T cells, combining CD3 with MEL-14 helps distinguish these subsets more precisely. The MEL-14 antibody is particularly useful when used together with antibodies to other cell surface markers to distinguish naïve, memory, and effector T cells.

Adhesion Molecules and Trafficking Markers

MEL-14 is frequently studied in conjunction with other adhesion molecules and trafficking markers. This makes sense given that CD62L itself is an adhesion molecule that mediates lymphocyte rolling on activated endothelium and homing to high endothelial venules (HEV) of peripheral lymphoid tissues. The protein binds to several glycosylated ligands including CD34, glycam-1, and MAdCam-1, which are important for understanding the complete picture of lymphocyte trafficking and adhesion.

Functional Context

The combination of MEL-14 with these various markers enables researchers to perform comprehensive analyses of immune cell subsets, activation states, and migration patterns. This multi-marker approach is essential in flow cytometry, immunohistochemistry, and functional blocking studies where understanding both cell identity and adhesion properties is crucial.

Key findings from scientific literature citing clone MEL-14 highlight its pivotal role in immunology as a monoclonal antibody specific for mouse L-selectin (CD62L), providing important insights into lymphocyte trafficking, cell adhesion mechanisms, and immune cell migration.

Essential findings include:

• Epitope Recognition: MEL-14 binds a specific epitope within the lectin domain of murine CD62L (the peripheral lymph node homing receptor), with detailed mapping assigning the critical region to the NH₂-terminal 53 amino acids of this domain.

• Conformational Dependence: The recognized epitope’s conformation may depend upon neighboring domains—recognition requires both lectin and EGF-like domains to be present, indicating that the lectin domain alone is insufficient for optimal antibody binding.

• Functional Blocking Activity: MEL-14 is a neutralizing antibody that can specifically and completely block lymphocyte binding to high endothelial venules (HEV) both in vitro and in vivo, resulting in effective inhibition of lymphocyte migration to lymph nodes.

• Mechanistic Implications: Blocking studies with MEL-14 established that the lectin domain of CD62L is directly involved in mediating lymphocyte adhesion to carbohydrate ligands on postcapillary venule endothelium, validating CD62L’s central role in lymphocyte homing and trafficking between the blood and lymphoid tissues.

• Cellular Expression: MEL-14 identifies CD62L expressed on most T and B lymphocytes, as well as neutrophils, monocytes, eosinophils, and some NK cell subsets.

• Research Utility: The MEL-14 clone is widely used in flow cytometry, immunohistochemistry, and functional assays to:

  • Assess and purify leukocyte subsets based on CD62L expression
  • Track lymphocyte recirculation and homing during inflammation
  • Model selectin function and leukocyte-endothelial interactions

• Impact on Understanding Inflammation: MEL-14, combined with functional studies (including those cited in classic immunological research), has been instrumental in exploring the molecular mechanisms of immune cell extravasation and trafficking—particularly during inflammatory responses.

In summary, MEL-14 citations have been foundational in demonstrating the importance of L-selectin in leukocyte homing, delineating the structural-functional relationship of its binding domains, and providing a key antibody tool for dissecting immune cell migration in vivo and in vitro.

The search results provided do not contain comprehensive information about how dosing regimens of clone MEL-14 vary across different mouse models. While several sources mention the MEL-14 antibody and its applications, specific dosing regimens across different mouse models are not detailed in the available search results.

Available Dosing Information

The search results primarily provide general usage guidelines rather than specific dosing regimens for different mouse models. For flow cytometry applications, the suggested concentration is ≤ 0.25 μg per 10⁶ cells in a volume of 100 μl. For western blotting, the recommended concentration is 1-10 μg/ml.

Functional Applications

The MEL-14 antibody has been documented in various functional studies. It has been shown to completely and specifically block binding of lymphocytes to high endothelial venules (HEV) in vitro and inhibit lymphocyte migration to lymph nodes in vivo. In delayed-type hypersensitivity (DTH) models, MEL-14 administration reduced ear swelling by blocking naive T cell recruitment to inflammation sites.

Research Considerations

The search results emphasize that titration of the reagent is recommended for optimal performance for each application, suggesting that dosing may need to be optimized based on the specific experimental context. One source mentions that each investigator should determine their own optimal working dilution for specific applications.

Without access to detailed protocols from published studies using MEL-14 across various mouse models (such as tumor models, inflammatory models, or different genetic backgrounds), a comprehensive comparison of dosing regimens cannot be provided based on the available information.