Human CLEC9a Antibody

(1 citations)   
  • 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 as a 0.2 µm filtered solution in PBS.
  • Endotoxin Level
    <0.10 EU per 1 μg of the antibody by the LAL method.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Binding Inhibition
    The binding of Recombinant Human CLEC9a Fc Chimera (Catalog # 6049-CL, 100 ng/mL) to a ligand expressed in Fas Ligand-treated A20 mouse B cell lymphoma cell line was maximally inhibited (>80%) by 10 µg/mL of the antibody, as detected by flow cytometry.

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.
Data Examples
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
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.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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:
    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.
  • 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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. The Effect of Chronic Hepatitis B Virus Infection on BDCA3+ Dendritic Cell Frequency and Function
    PLoS ONE, 2016;11(8):e0161235.
    Species: Human
    Sample Type: Whole Tissue
    Application: IHC - Paraffin embedded
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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Isotype Controls
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Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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