Mouse Mincle (CLEC4E) Antibody - Mouse IgG2a

Recombinant anti-mouse Mincle neutralizing and detection antibody

Neutralizing and detection antibody against mMincle

Anti-mMincle-mIgG2a is a monoclonal antibody (mAb) targeting mouse Mincle. This antibody was screened for neutralization activity and flow cytometry (see figure). 

Anti-mMincle-mIgG2a was previously extracted from hybridoma cells (clone 6G5). It is now expressed and produced in Chinese hamster ovary (CHO) cells, ensuring reliability and lot-to-lot reproducibility.

This antibody can be used together with HEK-Blue™ mMincle Cells for screening and neutralization assays to block Mincle signaling in response to stimulation by Mincle ligands.

Key features

• Each lot is functionally tested and validated.
• The complete sequence of the antibody construct has been verified.
• The absence of endotoxins is determined by the EndotoxDetect™ assay.

Macrophage-inducible C-type lectin (Mincle) is a C-type lectin receptor encoded by the C-type lectin domain family 4 member E (CLEC4E) gene. This receptor has emerged as an important player in innate immunity. It recognizes a variety of exogenous and endogenous stimuli, such as mycobacteria, some fungi, and necrotic cells [1, 2].

More details

The hybridoma-derived Anti-mMincle-IgG (mabg-mmcl-2) antibody has been replaced by Anti-mMincle-mIgG2a (mab10-mmcl), which is produced by recombinant technology and purified from CHO cells.

Read our review on C-Type Lectin Receptors

References:

1. Yamasaki S. et al., 2009. C-type lectin Mincle is an activating receptor for pathogenic fungus, Malassezia. PNAS 106(6): 1897–1902
2. Brown G.D., 2008. Sensing necrosis with Mincle. Nature Immunol. 9:1099-1100.

Figures

Cell surface staining of mouse Mincle. HEK-Blue mMincle cells were incubated with Anti-mMincle-mIgG2a or an isotype control for 1h at 4°C. Subsequently, a secondary PE‑labeled antibody was added and incubated at 4°C for 30 minutes. Cell surface staining was analyzed by flow cytometry.

Dose-dependent inhibition of HEK-Blue™ mMincle cell response using Anti-mMincle-mIgG2. HEK‑Blue™ mMincle cells were incubated for 1 hour with increasing concentrations of Anti-mMincle- mIgG2 (0.1 ng/ml - 10 μg/ml) prior to the addition of the Mincle agonist TDB (10 μg/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI‑Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Specifications

Application: Neutralization assay

Isotype: Mouse IgG2a, kappa

Recommended isotype control: Mouse IgG2a

Target: Mincle, CLEC4E

Species reactivity: Mouse

Clone: 6G5

Source: CHO cells

Production: Animal-free

Purification: Affinity chromatography

Physical form: Lyophilized

Formulation buffer: Sodium phosphate buffer with glycine, saccharose, and stabilizing agents

Preservative: Azide-free

Reconstitution buffer: Sterile water (not provided)

Purity: ≥ 95 %

Quality control: Each lot is functionally tested and validated.

Contents

Anti-mMincle-mIgG2a purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• mab10-mmcl: 100 µg
• mab10-mmcl-02: 2 x 100 µg

 Upon receipt, store lyophilized antibody at -20 °C.

 The lyophilized product is stable for at least 1 year.

 Avoid repeated freeze-thaw cycles.

Details

Mincle is a member of the C-type lectin receptor (CLR) family. Mincle recognizes a variety of exogenous and endogenous stimuli, such as mycobacteria, certain fungi, and necrotic cells [1, 2]. Exogenous ligands for Mincle include fungal α-mannose, and the mycobacterial glycolipid, trehalose-6’6’-dimycolate (TDM), also known as cord factor the immunostimulatory component of Mycobacterium tuberculosis [3].
Mincle also binds trehalose-6,6-dibehenate (TDB) which is a synthetic analog of TDM. Furthermore, Mincle senses damaged cells by recognizing endogenous damage-associated molecular patterns (DAMPs) [4]. Upon ligand recognition, Mincle interacts with the Fc receptor common γ-chain (FcRγ), which triggers intracellular signaling through Syk leading to CARD9-dependent NF-κB activation. Syk also induces the mobilization of intracellular calcium (Ca²⁺) and the activation of the calcineurin-NFAT pathway.

References:

1. Yamasaki S. et al., 2009​. C-type lectin Mincle is an activating receptor for pathogenic fungus, Malassezia. PNAS 106(6): 1897–1902. 
2. Brown GD. 2008.  Sensing necrosis with Mincle. Nature Immunol. 9:1099-1100. 
3. Ishikawa E. et al., 2009. Direct recognition of the mycobacterial glycolipid, trehalose dimycolate, by C-type lectin Mincle. J Exp Med. 206(13):2879-88.
4. Yamasaki S. et al., 2008. Mincle is an ITAM-coupled activating receptor that senses damaged cells. Nat Immunol. 9(10):1179-88.