Anti-Human CD226/DNAM-1 (LeoA1) ColorWheel^® Dye-Ready mAb

ColorWheel^® technology is a novel and proprietary method of creating your own antibody and dye combinations for use in flow cytometry. By incubating any ColorWheel^® antibody with any ColorWheel^® dye, researchers can quickly and simply produce primary conjugated antibodies for use in single color or multicolor flow cytometry analysis. Each ColorWheel^® antibody and dye is lyophilized for long-term storage, allowing for simplicity and flexibility without compromise. ColorWheel^® technology requires both ColorWheel^® antibody and ColorWheel^® dye for Flow Cytometry application.

CD226 antigen (UniProt: Q15762; also known as DNAX accessory molecule 1, DNAM-1, CD226) is encoded by the CD226 (also known as DNAM1) gene (Gene ID: 10666) in human. CD226 is a single-pass type I membrane protein of the immunoglobulin superfamily is synthesized with a signal peptide (aa 1-18), which is subsequently cleaved off to produce the mature protein that contains an extracellular domain (aa 19-254), a transmembrane domain (aa 255-275), and a cytoplasmic domain (aa 276-336). It also contains two Ig-like C2 type domains (aa 19-126 and 135-239). CD226 is expressed in T cells, NK cells, NK T cells, B cells, monocyte/macrophage, and dendritic cells. It is involved in cytotoxicity and cytokine secretion of T cells and NK cells and in platelet aggregation and activation. It serves as a receptor involved in intercellular adhesion and lymphocyte signaling. It promotes the migration, activation, proliferation, differentiation, and function of CD8 + T cells. CD155 and CD112 have been recognized as ligands for CD226. Solid tumor and hematological malignancies that express high levels of CD155 and CD112 are attractive targets for CTL attack via CD226-specific binding. CD226 expression is negatively correlated with the inhibitory function of Foxp3 + Tregs. TIGIT, a co-inhibitory molecule on T cells, is shown to exert immunosuppressive effects by competing with CD226 for the CD155 ligand. This ColorWheel^® Dye-Ready monoclonal antibody was developed using our Cat. No. MABT398; Anti-CD226/DNAM-1 monoclonal antibody, clone LeoA1. (Ref.: Huang, Z., et al. (2020). Front. Cell Dev. Biol. 8; 564; Xu, Z., et al. (2009). BMC Immunology 10; 34; Shibuya, A., et al. (1996). Immunity: 4(6); 573-581).

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Human T cells

Quality Control Testing

Evaluated by Flow Cytometry in Human peripheral blood mononuclear cells (PBMC).

Flow Cytometry Analysis: Staining of one million human peripheral blood mononuclear cells (PBMC) was performed with APC-conjugated anti-human CD3 and with 5 µL of a 1:1 mixture of Cat. No. CWA-1157, Anti-Human CD226/DNAM-1 (LeoA1) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye. Majority of CD49b expression was observed in CD3 positive population.

Tested Applications

Flow Cytometry Analysis: Staining of one million human peripheral blood mononuclear cells (PBMC) was performed with PE-conjugated anti-human CD3 and with 5 µL of a 1:1 mixture of Cat. No. CWA-1157, Anti-Human CD226/DNAM-1 (LeoA1) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye. Majority of CD49b expression was observed in CD3 positive population.

Flow Cytometry Analysis: Staining of one million fixed and permeabilized CHO cells stably transfected with human was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1157, Anti-Human CD226/DNAM-1 (LeoA1) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye or an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-PE ColorWheel^® Antibody-Ready Phycoerythrin (PE) Dye.

Flow Cytometry Analysis: Staining of one million fixed and permeabilized CHO cells stably transfected with human CD226 was performed using 5 µL of a 1:1 mixture of Cat. No. CWA-1157, Anti-Human CD226/DNAM-1 (LeoA1) ColorWheel^® Dye-Ready mAb and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye or a 1:1 mixture of an equivalent amount of Cat. No. CWA-1013 Mouse IgG1 isotype control and Cat. No. CWDS-APC ColorWheel^® Antibody-Ready Allophycocyanin (APC) Dye.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Clone LeoA1 is a mouse monoclonal antibody that detects CD226/DNAM-1.

Lyophilized from PBS containing D-Mannitol and Sucrose, normal appearance is a dried pellet. Reconstituted antibody solution is stable and functional as assessed by functional testing. Contains no biocide or preservatives, such as azide.

1.0 mg/mL after reconstitution at 25 µL in PBS. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Recommend storage of lyophilized product at 2-8°C. Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial. Reconstitute each vial by adding 25 µL PBS. Protect from light.

ColorWheel is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.