Dectin-1/CLEC7A Antibody [Unconjugated]

Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

Binding of cHis-sDectin-1 to Saccharomyces cerevisiae cells.Purified cHis-sDectin-1 was diluted to a concentration of 25 µg/ml in blocking buffer with 500 µg/ml, 100 µg/ml or no laminarin. This was added to Saccharomyces cerevisiae yeast cells from overnight culture in blocking buffer (PBS with 3% FBS). The cells were then probed with an mDectin-1 goat polyclonal antibody (1∶200; AF1756; R&D Systems) and AlexaFluor546-conjugated anti-goat antibody (1∶100; Catalog Number A-11056; Molecular Probes). After which, the cells were washed with PBS and fixed using 4% paraformaldehyde. The cells were then resuspended in PBS and visualized by phase contrast and fluorescent microscopy at (A) 200× magnification and (B) 600× magnification. Images are cropped or scaled to fit the illustration. cHis-sDectin-1 stained yeast cells was also analyzed by flow cytometry (C). Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

MTX amplification and characterization of sDectin-1 cell pools.(A) Western blotting of supernatant samples from cultures adapted to different MTX concentrations using an mDectin-1 goat polyclonal antibody (1∶1000; AF1756; R&D Systems) with a HRP conjugated anti-goat antibody (1∶2000; Catalog number V8051; Promega). cHis pool and nHis pool are cell pools producing sDectin-1 with histidine tagged at the C- and N-terminals respectively. (B) Shake flask batch culture cell growth and sDectin-1 production profiles of sDectin-1 producing cell pools. Values shown represent mean values obtained from three replicate flasks. Error bars indicate the standard deviation of the experiment. Image collected and cropped by CiteAb from the following publication (//dx.plos.org/10.1371/journal.pone.0052785), licensed under a CC-BY license. Not internally tested by R&D Systems.

OatN beta -glucan binds to HPCA, promotes microglia up-take of oatN and inhibits dectin-1 activation, whereas OatN digalactosyldiacylglycerol (DGDG) inhibits oatN beta -glucan access to dectin-1. H) A representative western blot of p-Syk in BV2 cells treated with Eth (200 mm) or Eth plus OBG (20 mg mL−1). Results represent 1 out of 3 independent experiments. Band intensity was quantified and normalized to beta -actin and the results are presented between the panels. I) The surface plasmon resonance (SPR) assay was performed to determine OBG binding to dectin-1 . (Upper panel) BG (commercial beta -glucan, 100 ng mL−1), PBLP (liposomes made by peripheral blood cell derived lipids), OatN (20 mg mL−1), and OBG (20 mg mL−1) were used as analytes. (Bottom panel) OBG-DGDG (OBG remove DGDG, 20 or 60 mg mL−1) and OBG-PC (OBG remove PC, 20 mg mL−1) were used as analytes. Results represent 1 out of 3 independent experiments. J) Dectin-1 binding to HPCA-Rab11a complex was detected by immunoprecipitation. HPCA complex in BV2 cells treated with Eth or OBG was pulled down by anti-HPCA antibody (1 mg mL−1). Dectin-1 and Rab11a were probed in a western blot. Results represent 1 out of 3 independent experiments. Band intensity was quantified and normalized by HPCA, and the results are presented between the panels. K) Confocal images of dectin-1 (green) overlay with Rab11a (red) in BV2 cells treated with Eth (200 mm) or Eth plus OBG (20 mg mL−1). Results represent 1 out of 3 independent experiments. Five random fields were photographed, and representative results are shown. Scale bars, 20 μm. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/34897972), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Mu
• Applications: WB, Flow, B/N, CyTOF-ready
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

Dectin-1/CLEC7A Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse Dectin-1/CLEC7A
  Phe69-Leu244
  Accession # Q6QLQ4
• Specificity: Detects mouse Dectin-1 in direct ELISAs and Western blots. In direct ELISAs and Western blots, less than 5% cross-reactivity with recombinant human Dectin-1 and recombinant mouse Dectin-2 alpha is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: CLEC7A
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Blockade of Receptor-ligand Interaction
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Western Blot 0.1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Dectin-1/CLEC7A Antibody [Unconjugated]

• Beta-glucan receptor
• BGR
• CD369
• CLEC7A
• CLECSF12
• CLECSF12DC-associated C-type lectin 1
• C-type (calcium dependent, carbohydrate-recognition domain) lectin, superfamilymember 12
• C-type lectin domain family 7 member A
• C-type lectin domain family 7, member A
• C-type lectin superfamily member 12
• Dectin1
• Dectin-1
• DECTIN1CANDF4
• Dendritic cell-associated C-type lectin 1
• dendritic cell-associated C-type lectin-1
• hDectin-1
• lectin-like receptor 1

Background

Dectin-1, also known as CLEC7A and the beta -glucan receptor, is a 43 kDa type II transmembrane C-type lectin that functions in the innate immune response to fungal pathogens. Although Dectin-1 resembles other CLEC molecules structurally, it binds ligands in a calcium-independent manner (1, 2). Mature mouse Dectin-1 is a 244 amino acid (aa) glycoprotein that consists of a short ITAM-containing cytoplasmic tail, a transmembrane segment, and a stalk and carbohydrate recognition domain (CRD) in the extracellular domain (3). The CRD of mouse Dectin-1 shares 61%, 60%, and 87% aa sequence identity with that of bovine, human, and rat Dectin-1, respectively. It shares 25-34% aa sequence identity with the CRD of other subgroup members CLEC-1, CLEC-2, CLEC9A, CLEC12B, LOX-1, and MICL. Mouse Dectin-1 is alternately spliced, generating a variant that lacks the stalk region (4). Mouse Dectin-1 is expressed on monocytes, macrophages, and neutrophils, and on some populations of dendritic cells and T cells (5). It is upregulated on macrophages by GM-CSF, IL-4, or IL-13 and downregulated by dexamethasone, IL-10, or LPS (6). The CRD selectively binds beta -glucan polymers, a major component of yeast and mycobacterial cell walls (7). Yeast beta -glucan is accessible to Dectin-1 only at sites of cell budding, and Dectin-1 does not recognize the filamentous form of yeast (8). Dectin-1 mediates the phagocytosis of zymosan particles and intact yeast (8-10). It co-localizes with TLR2 in the presence of zymosan, and the two receptors cooperate in ligand recognition and the propagation of proinflammatory signaling (9, 11-13). Dectin-1 interaction with the tetraspanin CD37 increases its stability on the cell membrane and inhibits ligand-induced signaling (14). Genetic knockout of Dectin-1 in mice increases their susceptibility to pathogenic infection (15, 16).

1. Kanazawa, N. (2007) J. Dermatol. Sci. 45:77.
2. Brown, G.D. (2006) Nat. Rev. Immunol. 6:33.
3. Ariizumi, K. et al. (2000) J. Biol. Chem. 275:20157.
4. Heinsbroek, S.E.M. et al. (2006) J. Immunol. 176:5513.
5. Taylor, P.R. et al. (2002) J. Immunol. 169:3876. 
6. Willment, J.A. et al. (2003) J. Immunol. 171:4569. 
7. Palma, A.S. et al. (2006) J. Biol. Chem. 281:5771.
8. Gantner, B.N. et al. (2005) EMBO J. 24:1277.
9. Gantner, B.N. et al. (2003) J. Exp. Med. 197:1107.
10. Kennedy, A.D. et al. (2007) Eur. J. Immunol. 37:467.
11. Brown, G.D. et al. (2003) J. Exp. Med. 197:1119.
12. Yadav, M. and J.S. Schorey (2006) Blood 108:3168.
13. Suram, S. et al. (2006) J. Biol. Chem. 281:5506.
14. Meyer-Wentrup, F. et al. (2007) J. Immunol. 178:154.
15. Saijo, S. et al. (2007) Nat. Immunol. 8:39.
16. Taylor, P.R. et al. (2007) Nat. Immunol. 8:31.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Dectin-1/CLEC7A Antibody (AF1756)(10)

Publications using AF1756

• Marie Goldmann, Franziska Schmidt, Zoltán Cseresnyés, Thomas Orasch, Susanne Jahreis, Susann Hartung et al. The Lipid Raft-Associated Protein Stomatin Is Required for Accumulation of Dectin-1 in the Phagosomal Membrane and for Full Activity of Macrophages against Aspergillus fumigatus mSphere 2023-02-21 [PMID: 36719247]
• Angela Yu-Huey Hsu, Sung-Tsang Hsieh Role of Dectin-1 in peripheral nerve injury Frontiers in Cellular Neuroscience 2022-07-28 [PMID: 35966205]
• Fangyi Xu, Jingyao Mu, Yun Teng, Xiangcheng Zhang, Kumaran Sundaram, Mukesh K. Sriwastva et al. Restoring Oat Nanoparticles Mediated Brain Memory Function of Mice Fed Alcohol by Sorting Inflammatory Dectin‐1 Complex Into Microglial Exosomes Small 2022-02-01 [PMID: 34897972]
• Georg Werner, Markus Damme, Martin Schludi, Johannes Gnörich, Karin Wind, Katrin Fellerer et al. Loss of TMEM 106B potentiates lysosomal and FTLD ‐like pathology in progranulin‐deficient mice EMBO reports 2020-10-05 [PMID: 32929860]
• JK Götzl, M Brendel, G Werner, S Parhizkar, L Sebastian, G Kleinberge, AV Colombo, M Deussing, M Wagner, J Winkelmann, J Diehl-Schm, J Levin, K Fellerer, A Reifschnei, S Bultmann, P Bartenstei, A Rominger, S Tahirovic, ST Smith, C Madore, O Butovsky, A Capell, C Haass Opposite microglial activation stages upon loss of PGRN or TREM2 result in reduced cerebral glucose metabolism EMBO Mol Med, 2019-06-01;0(0):. 2019-06-01 [PMID: 31122931] (Western Blot, Mouse)
• JAT de Albuque, PP Banerjee, A Castoldi, R Ma, NB Zurro, LH Ynoue, C Arslanian, MUW Barbosa-Ca, JEM Correia-De, FG Weiler, MR Dias-da-Si, M Lazaretti-, LA Pedroza, NOS Câmara, E Mace, JS Orange, A Condino-Ne The Role of AIRE in the Immunity Against Candida Albicans in a Model of Human Macrophages Front Immunol, 2018-03-21;9(0):567. 3/21/2018 [PMID: 29666621]
• Yamasaki T, Ariyoshi W, Okinaga T, Adachi Y, Hosokawa R, Mochizuki S, Sakurai K, Nishihara T The dectin 1 agonist curdlan regulates osteoclastogenesis by inhibiting nuclear factor of activated T cells cytoplasmic 1 (NFATc1) through Syk kinase. J Biol Chem, 2014-05-12;289(27):19191-203. 2014-05-12 [PMID: 24821724] (Western Blot, Mouse)
• Say Kong Ng, Tessa Rui Min Tan, Yang Wang, Daniel Ng, Lin-Tang Goh, Muriel Bardor et al. Production of Functional Soluble Dectin-1 Glycoprotein Using an IRES-Linked Destabilized-Dihydrofolate Reductase Expression Vector PLoS ONE 2012-12-26 [PMID: 23300776]
• Fuller GL, Williams JA, Tomlinson MG, Eble JA, Hanna SL, Pohlmann S, Suzuki-Inoue K, Ozaki Y, Watson SP, Pearce AC The C-type lectin receptors CLEC-2 and Dectin-1, but not DC-SIGN, signal via a novel YXXL-dependent signaling cascade. J. Biol. Chem., 2007-03-05;282(17):12397-409. 2007-03-05 [PMID: 17339324] (Neutralization, Mouse)
• Rappleye CA, Eissenberg LG, Goldman WE Histoplasma capsulatum alpha-(1,3)-glucan blocks innate immune recognition by the beta-glucan receptor. Proc. Natl. Acad. Sci. U.S.A., 2007-01-16;104(4):1366-70. 2007-01-16 [PMID: 17227865] (ICC, Mouse)

Bioinformatics

• Gene Symbol: CLEC7A
• Uniprot:
  • Q6QLQ4()