Detection of DNAM-1/CD226 in Mouse Splenoctyes by Flow Cytometry.
Mouse splenocytes were stained with Rat Anti-Mouse CD8 alpha PE‑conjugated Monoclonal Antibody (Catalog # FAB116P) and either (A) Rat Anti-Mouse DNAM-1/CD226 APC‑conjugated Monoclonal Antibody (Catalog # FAB4436A) or (B) Rat IgG2B Allophycocyanin Isotype Control (Catalog # IC013A). View our protocol for Staining Membrane-associated Proteins.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
12 months from date of receipt, 2 to 8 °C as supplied.
DNAX accessory molecule-1 (DNAM-1), also known as CD226, is a 63-65 kDa type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily. Mature mouse DNAM has a 236 amino acid (aa) extracellular domain (ECD) that contains two Ig-like C2-set domains, and possesses a 58 aa cytoplasmic region that contains motifs for binding PDZ domains and band 4.1 family proteins. Within the ECD, mouse DNAM-1 shares 52% and 86% aa sequence identity with human and rat DNAM-1, respectively. Additional cDNA transcripts of mouse DNAM-1 may give rise to secreted or transmembrane isoforms with ECD deletions. DNAM-1 is expressed on several lymphoid and myeloid cell types including NK cells, CD8+ T cells, CD4+ Th1 cells, NKT cells, monocytes, mast cells and plateletes and interacts with CD155/PVR and Nectin-2/CD112 in trans, and with LFA-1 in cis. Ligation of DNAM-1 promotes the activation of NK cells, CD8+ T cells, and mast cells, induces dendritic cell maturation, initiates megakaryocyte and activated platelet adhesion to vascular endothelial cells, and stimulates monocyte extravasation; conversely, it inhibits the formation of osteoclasts. Platelet-endothelium interactions that are mediated by DNAM-1 enable the metastasis of tumor cells to the lung. On activated, but not resting NK, T, and mast cells, the cis association of DNAM-1 with CD18 contributes to tyrosine and serine phosphorylation of DNAM-1 during activation. Finally, DNAM-1 is now known to differentiate between DNAM-1+ NK cells that produce inflammatory cytokines, an DNAM-1- NK cells that secrete MIP-1 chemokines.
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