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

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

Product No.: C6118

[product_table name="All Top" skus="C2118"]

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Clone
MEL-14
Target
CD62L
Formats AvailableView All
Product Type
Monoclonal Antibody
Alternate Names
L-selectin, LECAM-1, Ly-22, LAM-1, MEL-14
Isotype
Rat IgG2a κ
Applications
B
,
CyTOF®
,
Depletion
,
FA
,
FC
,
IHC FF
,
in vivo
,
IP
,
WB

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Select Product Size

Data

Anti-Mouse CD62L CyTOF™ Data
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Antibody Details

Product Details

Reactive Species
Mouse
Host Species
Rat
Recommended Isotype Controls
Recommended Dilution Buffer
Immunogen
C3H/eb mouse B lymphoma 38C-13
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
<0.5 EU/mg as determined by the LAL method
Purity
≥98% monomer by analytical SEC
>95% by SDS Page
Formulation
This monoclonal antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Functional grade preclinical antibodies are manufactured in an animal free facility using in vitro cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s Purified Functional PLATINUM™ antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Country of Origin
USA
Shipping
Next Day 2-8°C
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for this MEL-14 antibody for staining cells in flow cytometry is ≤ 0.25 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
WB The suggested concentration for this MEL-14 antibody for use in western blotting is 1-10 μg/ml.
Additional Applications Reported In Literature ?
CyTOF®
IHC FF
IP
B
Additional Reported Applications For Relevant Conjugates ?
IHC (Paraffin)
For specific conjugates of this clone, review literature for suggested application details.
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
Rat Anti-Mouse CD62L (Clone MEL-14) recognizes an epitope on Mouse CD62L. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.
Background
CD62L is a 74-95 kD glycoprotein is a member of the selectin family and is expressed on the majority of B and naïve T cells, a subset of memory T cells, monocytes, granulocytes, most thymocytes, and a subset of NK cells. Furthermore, CD62L is a cell adhesion molecule that binds to many glycoprotein ligands including CD34, GlyCAM-1, and PSGL-1. Clone MEL-14 is reported as a Nutralization antibody.
Antigen Distribution
Subsets of B and T cells, monocytes, granulocytes, subset of NK cells
Ligand/Receptor
CD34, GlyCAM-1, MAdCAM-1
Function
Lymphocyte homing to HEV, rolling on activated endothelium
PubMed
NCBI Gene Bank ID
Research Area
Cell Adhesion
.
Cell Biology
.
Costimulatory Molecules
.
Immunology
.
Innate Immunity

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Clone MEL-14 is a rat monoclonal antibody targeting mouse CD62L (L-selectin) and is used in vivo mouse studies primarily for two applications: functional blockade of lymphocyte homing to lymph nodes and as a surface marker for flow cytometric identification of lymphocyte subpopulations.

Key uses in in vivo mouse studies:

  • Blocking Lymphocyte Homing: MEL-14 can be administered to mice to block the interaction between lymphocytes and high endothelial venules (HEV) in lymph nodes, thereby inhibiting lymphocyte migration or homing to lymph nodes. Pre-incubation of lymphocytes with MEL-14 or systemic administration in mice completely and specifically blocks binding of lymphocytes to HEV in vitro and their migration to lymph nodes in vivo. This property is often exploited in studies analyzing immune cell trafficking, lymphocyte localization, and immune response dynamics.

  • Functional Depletion: By blocking CD62L, MEL-14 can functionally deplete naive T cells from entering lymph nodes, which allows researchers to assess the role of lymphocyte trafficking in immunity or inflammation models.

  • Phenotyping by Flow Cytometry: Though much of the cited use is in vitro, MEL-14 is also widely used ex vivo (on lymphocytes from mice, sometimes after in vivo exposure to treatments) for flow cytometric identification of naive (CD62L^hi) versus activated/memory (CD62L^lo) T cell populations. Flow cytometry using MEL-14 facilitates the detailed study of T cell subsets, their activation status, and trafficking capacity.

Important considerations:

  • Functional Assays: When using MEL-14 in functional blockade studies, sodium azide should be removed from antibody preparations, as it can be toxic in vivo.
  • Specificity: MEL-14 binding is specific for mouse CD62L and does not cross-react with other selectins.
  • Expression: CD62L is expressed on most T and B lymphocytes, neutrophils, monocytes, and eosinophils, meaning MEL-14 can affect multiple immune cell types.

Summary Table: MEL-14 Uses in In Vivo Mouse Experiments

ApplicationMechanism/Utility
Homing blockadeInhibits lymphocyte migration to LN by blocking CD62L
Functional depletionPrevents naive T cells from entering lymphoid organs
Phenotyping (ex vivo)Distinguishes naive vs. activated/memory T cells

In summary, MEL-14 is chiefly used in vivo to block lymphocyte trafficking to lymph nodes, allowing researchers to dissect the role of cellular localization in immune responses, as well as to characterize lymphocyte phenotypes ex vivo after in vivo manipulations.

The correct storage temperature for the sterile packaged clone MEL-14 antibody is 2–8°C (typically a standard refrigerator temperature), stored undiluted and protected from prolonged exposure to light. Do not freeze the antibody.

Additional details:

  • This temperature range is specific to MEL-14 antibodies as listed by multiple suppliers and applies to both labeled and unlabeled forms.
  • Prolonged exposure to light should be avoided to prevent potential degradation, especially for conjugated antibodies.
  • General sterile storage recommendations for other types of sterile supplies (not antibodies) may suggest higher temperatures (up to 18–24°C), but antibody reagents like MEL-14 require 2–8°C as specified by manufacturers.

Commonly used antibodies or proteins frequently reported in the literature alongside MEL-14 (anti-mouse CD62L/L-selectin) include markers of T and B cell subsets and activation, as well as additional adhesion molecules and trafficking markers.

Frequently used antibodies/proteins with MEL-14:

  • CD3 (pan-T cell marker)
  • CD4, CD8 (T helper and cytotoxic T cell markers)
  • CD19, B220 (B cell markers)
  • CD44 (activation/memory marker, also involved in cell adhesion/migration)
  • CD62P (P-selectin) and CD62E (E-selectin): other selectin family adhesion molecules
  • CD11a, CD11b, CD18 (integrins: LFA-1, Mac-1-related): leukocyte adhesion and trafficking
  • CD34, GlyCAM-1, PSGL-1: endothelial or ligand molecules relevant to leukocyte trafficking and often studied in functional or co-staining experiments
  • CCR7, CXCR4: chemokine receptors important for lymphocyte migration and homing

Context and experimental rationale:

  • MEL-14 is routinely used to distinguish naïve T and B lymphocytes (high CD62L) from effector/memory (low CD62L) cells in multi-color flow cytometry panels.
  • CD62L is also examined in conjunction with other homing and activation markers (e.g., CD44) to define lymphocyte differentiation and migration status.
  • In studies on lymphocyte trafficking or tissue residency, MEL-14 may be paired with antibodies to molecules like CD34, GlyCAM-1, or integrins, to study endothelial interactions and the multi-step cascade of lymphocyte migration.

Experimental protocols and applications:

  • Flow cytometry panels often include MEL-14 with CD3, CD4, CD8, B220, CD44, and CD11a/CD18 for immunophenotyping.
  • Blocking/neutralization experiments targeting CD62L (MEL-14) may use paired controls or isotypes, and sometimes co-blockade with anti-CD44 or anti-integrin antibodies, especially in vivo studies of lymphocyte migration or trafficking.
  • For immunoprecipitation or Western blot, additional proteins such as E-selectin, P-selectin, or chemokine receptors may also be included, depending on the migration axis under investigation.

Alternative nomenclature: In the literature, CD62L may also be referenced as L-selectin, LECAM-1, Ly-22, LAM-1, or lymphocyte homing receptor.

In sum, MEL-14 is most often used together with markers for lymphocyte subset identification, activation (e.g., CD44), and other trafficking-related proteins (e.g., integrins, selectins, chemokine receptors) when studying lymphocyte biology and homing in mice.

The MEL-14 clone has generated significant findings in scientific literature, particularly regarding its role in understanding lymphocyte trafficking and selectin function. This rat anti-mouse CD62L monoclonal antibody has been instrumental in advancing our knowledge of cell adhesion mechanisms and immune cell migration.

Recognition and Binding Properties

MEL-14 recognizes mouse L-selectin (CD62L), an adhesion molecule expressed on most T and B lymphocytes, neutrophils, monocytes, and eosinophils. The antibody specifically binds to an epitope within the lectin domain of the murine peripheral lymph node homing receptor (pln HR). Through detailed epitope mapping studies, researchers determined that the MEL-14 recognition site is located within the NH2-terminal 53 amino acids of the lectin domain.

Functional Blocking Capabilities

One of the most significant findings is MEL-14's ability to completely and specifically block lymphocyte-endothelial interactions. Pre-incubation of lymphocytes with MEL-14 completely blocks binding of lymphocytes to high endothelial venules (HEV) in vitro and prevents the migration of lymphocytes to lymph nodes in vivo. This blocking activity demonstrates that MEL-14 interferes with the same cellular interactions that are inhibited by PPME, a yeast cell wall polyphosphomannan carbohydrate.

Structural Insights and Domain Dependencies

Research using MEL-14 revealed important structural requirements for proper epitope recognition. While the antibody recognized truncated homing receptor constructs containing both the lectin and EGF domains, antibody recognition was lost when the lectin domain alone was expressed. This finding suggests that the MEL-14 epitope's conformation may be dependent upon the presence of the EGF domain, highlighting the importance of proper protein folding and domain interactions for antibody recognition.

Mechanistic Understanding of Lymphocyte Trafficking

The functional blocking capabilities of MEL-14 have provided crucial evidence supporting the hypothesis that the peripheral lymph node homing receptor's lectin domain is directly involved in binding lymphocytes to carbohydrate ligands on peripheral postcapillary venule endothelium. This finding has been fundamental to understanding the molecular mechanisms underlying lymphocyte homing and trafficking between blood circulation and lymphoid tissues.

Technical Applications and Research Impact

MEL-14 has proven valuable across multiple research applications, including flow cytometry, immunohistochemistry, and functional assays. Its ability to efficiently block lymphocyte-endothelial interactions has made it an essential tool for studying immune cell migration patterns and understanding the role of selectins in inflammatory processes. The antibody's specificity and blocking function have established it as a standard reagent for investigating L-selectin-mediated cellular interactions in both basic research and applied studies of immune system function.

References & Citations

B
CyTOF®
Depletion
FA
Flow Cytometry
IHC FF
in vivo Protocol
Immunoprecipitation Protocol
General Western Blot Protocol

Certificate of Analysis

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.