Anti-human Dectin-1 (CLEC7A) antibody

Recombinant anti-human Dectin-1 neutralizing and detection antibody

Neutralizing and detection antibody against hDectin-1

Anti-hDectin-1-mIgG1 is a monoclonal antibody (mAb) targeting human Dectin-1. This antibody was screened for neutralization activity and flow cytometry (see figures).

Anti-hDectin-1-mIgG1 was previously extracted from hybridoma cells (clone 22H8). It is now expressed and produced in Chinese hamster ovary (CHO) cells, ensuring reliability and lot-to-lot reproducibility. Anti-hDectin-1-mIgG1 recognizes hDectin-1 isoforms A and B. 

This antibody can be used together with HEK-Blue™ hDectin-1b Cells for screening and neutralization assays to block hDectin-1 signaling in response to stimulation by Dectin 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.

Dendritic cell-associated C-type lectin-1 (Dectin-1) is a C-type lectin receptor (CLR) that recognizes β-glucans, glucose polymers found in the cell walls of fungi, and thus plays an important role in antifungal innate immunity. Dectin-1  is encoded by the C-type lectin domain family 7 member A (CLEC7A) gene and alternatively spliced into 2 major isoforms: a full-length A isoform and a ‘stalkless’ B isoform, which do not induce the same response to soluble and particulate β-glucans [1, 2].

More details

The hybridoma-derived Anti-hDectin-1-IgG (mabg-hdect-2) antibody has been replaced by Anti-hDectin-1-mIgG1 (mab9-hdect1), which is produced by recombinant technology and purified from CHO cells.

Read our review on Dectin-1 & β-glucans.

Read our review on Dectin-1 & inflammasomes.

References:

1. Brown G.D. et al., 2003. Dectin-1 mediates the biological effects of beta-glucans. J Exp Med. 197: 1119- 24.
2. Heinsbroek S.E. et al., 2006. C-type Lectin Receptor: Old Friend and New Player. Med Chem. 2017;13(6):536-543.

Figures

Cell surface staining of human Dectin-1. HEK-Blue™ hDectin-1b cells were incubated with Anti-hDectin-1-mIgG1 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™ hDectin-1b cell response using Anti-hDectin-1-mIg1. HEK-Blue™ hDectin-1b cells were incubated for 1 hour with increasing concentrations of Anti-hDectin-1-mIgG1 (0.1 ng/ml - 10 μg/ml) prior to the addition of the Dectin agonist Heat-killed S. cerevisiae (1 x 107 cells). 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 IgG1, kappa

Recommended isotype control: Mouse IgG1

Target: Dectin-1 (CLEC7A)

Species reactivity: Human

Clone: 22H8

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-hDectin-1-mIgG1 purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• mab9-hdect1: 100 µg
• mab9-hdect1​-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

Dectin-1 is a member of the C-type lectin receptor (CLR) family and plays an important role in antifungal innate immunity. In humans and mice, it is alternatively spliced into two major isoforms, a full-length A isoform and a ‘stalkless’ B isoform [1, 2]. It is expressed on phagocytic cells, including macrophages and neutrophils.
Dectin-1 is a specific receptor for β-glucans [2]. β-Glucans are glucose polymers found in the cell wall of fungi, including the yeasts Saccharomyces cerevisiae and Candida albicans. Upon binding to its ligand, Dectin-1 triggers phagocytosis and signaling through the kinase Syk and the adaptors CARD9-Bcl10-Malt1 leading to the production of reactive oxygen species (ROS), activation of NF-κB and subsequent production of pro-inflammatory cytokines [3].
Notably, Dectin-1 signaling has been shown to collaborate with Toll‑like receptor 2 (TLR2) signaling to enhance the responses triggered by each receptor [4].

1. Fischer M. et al., 2017. Isoform localization of Dectin-1 regulates the signaling quality of anti-fungal immunity. Eur J Immunol. 47(5):848-859.
2. Hou H. et al., 2017. C-type Lectin Receptor: Old Friend and New Player. Med Chem. 13(6):536-543.
3. Drummond RA. & Lionakis MS., 2016. Mechanistic Insights into the Role of C-Type Lectin Receptor/ CARD9 Signaling in Human Antifungal Immunity. Front Cell Infect Microbiol. 6:39.
4. Romero MM. et al., 2016. Reactive oxygen species production by human dendritic cells involves TLR2 and dectin-1 and is essential for efficient immune response against Mycobacteria. Cell Microbiol. 18(6):875-86.