CLEC9a Antibody

Western blot shows lysates of human CD14+ peripheral blood mononuclear cells (PBMC). PVDF Membrane was probed with 1 µg/mL of Sheep Anti-Human CLEC9a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6049) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for CLEC9a at approximately 45 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.

CLEC9a was detected in immersion fixed paraffin-embedded sections of human spleen using Sheep Anti-Human CLEC9a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6049) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu
• Applications: WB, IHC, B/N
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

CLEC9a Antibody Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human CLEC9a
  Lys57-Val241
  Accession # Q6UXN8
• Specificity: Detects human CLEC9a in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant mouse CLEC9a is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: CLEC9A
• Purity: Immunogen affinity purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • Immunohistochemistry 5-15 ug/mL
  • Binding Inhibition 10 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 as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Purity: Immunogen affinity purified
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.2 mg/mL.

Notes

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

Alternate Names for CLEC9a Antibody

• CD370
• CLEC9a
• C-type lectin domain family 9 member A
• C-type lectin domain family 9, member A
• DNGR1
• HEEE9341
• UNQ9341

Background

CLEC9a (C-type lectin domain family 9 member A), also known as DNGR-1, is a type II transmembrane glycoprotein member of the C-type lectin superfamily (1, 2). Mature human CLEC9a consists of a 35 amino acid (aa) cytoplasmic domain with an ITAM-like motif, a 21 aa transmembrane segment, and a 185 extracellular domain (ECD) that contains a stalk region and one C-type lectin domain (CTLD) (3‑5). Within the ECD, human CLEC9a shares 57% aa sequence identity with mouse and rat CLEC9a. Although the CTLD of CLEC9a structurally resembles that of other C-type lectins, it lacks the conserved residues that typically mediate calcium and carbohydrate binding. CLEC9a is expressed as a disulfide-linked homodimer of approximately 45-50 kDa N-glycosylated subunits (3, 5). Human CLEC9a expression is restricted to a subpopulation of BDCA-3⁺ conventional dendritic cells (cDC) and CD16^- monocytes (3‑5). BDCA-3⁺ cDC are analagous to mouse CD8⁺ cDC which are specialized in antigenic cross-presentation in complex with MHC class I molecules (6). In mouse, CLEC9a is additionally expressed on plasmacytoid dendritic cells (4, 5). CLEC9a ligation enhances antigen uptake and processing, leading to presentation on MHC class I and cytotoxic T cell (CTL) priming (3‑5). In mouse, CLEC9a recognizes normally intracellular determinant(s) of necrotic cells and mediates their uptake by the dendritic cell (7). The subsequent antigenic cross-presentation to CTL is important for clearing necrotic cellular debris (7). CLEC9a signaling triggers activation of the tyrosine kinase Syk (3, 7).

1. Huysamen, C. and G.D. Brown (2009) FEMS Microbiol. Lett. 290:121.
2. Geijtenbeek, T.B.H. et al. (2004) Annu. Rev. Immunol. 22:33.
3. Huysamen, C. et al. (2008) J. Biol. Chem. 283:16693.
4. Caminschi, I. et al. (2008) Blood 112:3264.
5. Sancho, D. et al. (2008) J. Clin. Invest. 118:2098.
6. Dudziak, D. et al. (2007) Science 315:107.
7. Sancho, D. et al. (2009) Nature 458:899.

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 CLEC9a Antibody (AF6049)(1)

Publication using AF6049

• The Effect of Chronic Hepatitis B Virus Infection on BDCA3+ Dendritic Cell Frequency and Function PLoS ONE, 2016;11(8):e0161235. 2016 [PMID: 27529176] (IHC-P, Human)

Bioinformatics

• Gene Symbol: CLEC9A
• Uniprot:
  • Q6UXN8()