Human CLEC9a Antibody

Catalog #: AF6049
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6049
AF6049-SP

Detection of Human CLEC9a by Western Blot.

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All CLEC9a Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CLEC9a Antibody Summary

Species Reactivity

Human

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

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

Mouse myeloma cell line NS0-derived recombinant human CLEC9a
Lys57-Val241
Accession # Q6UXN8

Formulation

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.

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human CLEC9a by Western Blot. 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.

Immunohistochemistry

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CLEC9a in Human Spleen. 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.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Background: CLEC9a

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).

References

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

Long Name

C-type Lectin-like Receptor 9

Entrez Gene IDs

283420 (Human); 232414 (Mouse)

Alternate Names

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

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Citation for Human CLEC9a Antibody

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