|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.|
|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.|
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).
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