Human peripheral blood mononuclear cells (PBMCs) gated on CD3^-cells were stained with Mouse Anti-Human NCAM-1/CD56 APC-conjugated Monoclonal Antibody (Catalog # FAB2408A) and either (A) Mouse Anti-Human DNAM-1/CD226 Monoclonal Antibody (Catalog # MAB666) or (B) Mouse IgG₁Isotype Control (Catalog # MAB002) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B).

Neutralization of DNAM-1/CD226 Fc Chimera by Human DNAM-1/CD226 Antibody

In a functional ELISA, Human DNAM-1/CD226 Antibody (Catalog # MAB666) blocks the binding of Recombinant Human DNAM-1/CD226 Fc Chimera (666-DN) to Biotinylated Recombinant Human CD155/PVR Fc Chimera Avi-tag (AVI9174). The Neutralization Dose (ND₅₀) for this effect is typically 0.300-6.00 µg/mL.

Applications for Human DNAM-1/CD226 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/10⁶ cells
Sample: Human peripheral blood mononuclear cells (PBMCs) gated on CD3^- cells

Western Blot

1 µg/mL
Sample: Recombinant Human DNAM-1/CD226 Fc Chimera (Catalog # 666-DN) under non-reducing conditions only

Neutralization

In a functional ELISA, Human DNAM-1/CD226 Antibody (Catalog # MAB666) blocks the binding of Recombinant Human DNAM-1/CD226 Fc Chimera (Catalog # 666-DN) to Biotinylated Recombinant Human CD155/PVR Fc Chimera Avi-tag (Catalog # AVI9174). The Neutralization Dose (ND₅₀) for this effect is typically 0.300-6.00 µg/mL.

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using MAB666 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from ascites

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

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: DNAM-1/CD226

DNAX accessory molecule-1 (DNAM-1), also known as CD226, is a 65 kDa type I transmembrane glycoprotein in the immunoglobulin superfamily (1). Mature human DNAM-1 contains a 236 amino acid (aa) extracellular domain (ECD) with two Ig-like C2-set domains and a 61 aa cytoplasmic region that contains motifs for binding PDZ domains and band 4.1 family proteins (1, 2). Within the ECD, human DNAM-1 shares 50% and 52% aa sequence identity with mouse and rat DNAM-1, respectively. DNAM-1 is expressed on multiple lymphoid and myeloid cell types and interacts with CD155/PVR and Nectin-2/CD112 (3, 4). Ligation of DNAM-1 promotes the activation of NK cells, CD8⁺ T cells, and mast cells (2‑6), dendritic cell maturation, megakaryocyte and activated platelet adhesion to vascular endothelial cells, and monocyte extravasation; it inhibits the formation of osteoclasts (7‑10). Platelet-endothelium interactions mediated by DNAM-1 enable the metastasis of tumor cells to the lung (11). In activated, but not in resting NK, T, and mast cells, the cis association of DNAM-1 with CD18 contributes to the tyrosine and serine phosphorylation of DNAM-1 during activation (6, 9, 12‑14).

References

Fuchs, A. and M. Colonna (2006) Semin. Cancer Biol. 16:359.
Shibuya, A. et al. (1996) Immunity 4:573.
Bottino, C. et al. (2003) J. Exp. Med. 198:557.
Tahara-Hanaoka, S. et al. (2004) Int. Immunol. 16:533.
Dardalhon, V. et al. (2005) J. Immunol. 175:1558.
Bachelet, I. et al. (2006) J. Biol. Chem. 281:27190.
Reymond, N. et al. (2004) J. Exp. Med. 199:1331.
Kakehi, S. et al. (2007) Mol. Cell. Biochem. 301:209.
Kojima, H. et al. (2003) J. Biol. Chem. 278:36748.
Tahara-Hanaoka, S. et al. (2006) Blood 107:1491.
Morimoto, K. et al. (2007) Oncogene July 16 epub.
Shibuya, K. et al. (1999) Immunity 11:615.
Shibuya, K. et al. (2003) J. Exp. Med. 198:1829.
Shibuya, A. et al. (1998) J. Immunol. 166:1671.

Long Name

DNAX Accessory Molecule 1

Alternate Names

CD226, DNAM1, PTA1, TLiSA1

Entrez Gene IDs

10666 (Human); 225825 (Mouse); 307199 (Rat); 102117316 (Cynomolgus Monkey)

Gene Symbol

CD226

UniProt

Q15762

Additional DNAM-1/CD226 Products

Product Documents for Human DNAM-1/CD226 Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human DNAM-1/CD226 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry