Detection of DNAM-1/CD226 in Mouse Splenocytes by Flow Cytometry.
Mouse splenocytes were stained with Rat Anti-Mouse CD8 alpha APC-conjugated Monoclonal Antibody (Catalog # FAB116A) and either (A) Rat Anti-Mouse DNAM-1/CD226 Monoclonal Antibody (Catalog # MAB4436) or (B) Rat IgG2B Isotype Control (Catalog # MAB0061) followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B).
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
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.
DNAX accessory molecule-1 (DNAM-1), also known as CD226, is a 65 kDa type I transmembrane glycoprotein that belongs to the immunoglobulin superfamily (1). 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 (1, 2). 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 and interacts with CD155/PVR and Nectin-2/CD112 (2‑4). Ligation of DNAM-1 promotes the activation of NK cells, CD8+ T cells, and mast cells (3‑7), 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 (8‑11). Platelet-endothelium interactions that are mediated by DNAM-1 enable the metastasis of tumor cells to the lung (12). 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 (7, 10, 13‑15).
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