- Other Names
- DNAX accessory molecule-1, DNAM-1, CD226, Platelet and T-cell activation antigen 1, PTA1, T lineage specific activation antigen 1 antigen, TLiSA
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- Product Citations
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DNAM-1, also known as CD226, was first identified as T lineage-specific activation antigen (TLiSA) and subsequently renamed as Platelet and T-cell activation antigen 1 (PTA). DNAM-1 consists of two Ig-like domains on its extracellular portion, a transmembrane region and a cytoplasmic region containing binding motifs for members of band 4.1 family and membrane-associated guanylate kinase homolog (MAGUK) family. DNAM-1 is predominantly expressed on cell surface of T cells, NK cells, monocytes/ macrophages, platelets and megakaryocytes and a subset of B cells. DNAM-1 serves as a signaling-transducing adhesion molecule, and it physically and functionally associates with LFA-1 (CD11a/CD18) on NK and T cells. The clustering of LFA-1 induces Fyn-mediated phosphorylation of DNAM-1 and signaling. Several evidences demonstrated that DNAM-1 is involved in NK and T cell-mediated cytotoxicity (CTL) against ligand-expressing tumor cells. Two ligands for DNAM-1 have been identified, the poliovirus receptor (PVR/CD155) and its family member nectin-2 (PRR/CD112), which are located at cell junctions and broadly expressed on epithelial, endothelial and neuronal cells. Additionally, DNAM-1 regulates monocyte extravasation through PVR expressed by endothelial cells. It has reported that DNAM-1 is specifically expressed on differentiated Th1 cells, but not to Th2 or Th0 cells. Antibody blocking assay demonstrated that DNAM-1 may play a role in polarization of T cells, activation and effectors of Th1 cells. PVRIG (CD112R) is identified as a novel immune checkpoint to inhibit T cell response by competing DNAM-1 with CD112.Product Details
- Human DNAM-1/CD226, amino acid (Glu19-Asn247) (Accession: #Q15762.2), with a linker (GSSR), a C-terminal human IgG1 (Pro100-Lys330) and a 6x His tag, was expressed in 293E cells.
- Molecular Mass
- The 464 amino acid recombinant protein has a predicted molecular mass of approximately 52.3 kD. The DTT-reduced and non-reduced protein migrates at approximately 65 and 130 kD respectively by SDS-PAGE. The predicted N-terminal amino acid is Glu.
- > 95%, as determined by Coomassie stained SDS-PAGE.
- 0.22 µm filtered protein solution is in PBS, pH 7.2.
- Endotoxin Level
- Less than 0.1 EU per μg protein as determined by the LAL method.
- 10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg and larger sizes are lot-specific and bottled at the concentration indicated on the vial (please contact technical support for concentration, or use our Lookup tool if you have a lot number.)
- Storage & Handling
- Unopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
- ED50 = 0.25 – 1.0 μg/mL as measured by the ability of immobilized protein to support the adhesion of U937 cells. Calcein-AM (Cat. No. 425201) was used to measure the number of adherent cells.
- Application Notes
BioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are validated in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at firstname.lastname@example.org.
(PubMed link indicates BioLegend citation)
- Fuchs A, Colonna M. 2006. Semin. Cancer Biol. 16: 359.
- Kojima H, et al. 2003. J. Biol. Chem. 278: 36748.
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- Tahara-Hanaoka S, et al. 2006. Blood. 107: 1491.
- Bottino C, et al. 2003. J. Exp. Med. 198: 557.
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- Dardalhon V, et al. 2005. J. Immunol. 175: 1558.
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- Zhu Y, et al. 2016. J. Exp. Med. 213: 167.
- Disulfide bond-linked homodimer, Ig domain containing adhesion molecule.
Peripheral blood T cells, NK cells, monocytes/macrophages, platelets and megakaryocytes and a subset of B cells; plasma membrane.
- It is involved in intercellular adhesion, lymphocyte signaling, cytotoxicity and lymphokine secretion mediated by cytotoxic T-lymphocyte (CTL) and NK cells.
- Epithelial, endothelial and neuronal cells; LFA-1.
- PVR (CD155) and Nectin-2 (CD112)
- Measured by the ability of immobilized protein to support the adhesion of U937 cells.
- Biology Area
- Molecular Family
- Adhesion Molecules, CD Molecules, Soluble Receptors
- Gene ID
- 10666 View all products for this Gene ID
- View information about DNAM-1 on UniProt.org
- Does specific activity of a recombinant protein vary between lots?
Specific activity will vary for each lot and for the type of experiment that is done to validate it, but all passed lots will have activity within the established ED50 range for the product and we guarantee that our products will have lot-to-lot consistency. Please conduct an experiment-specific validation to find the optimal ED50 for your system.
- Have your recombinants been tested for stability?
Our testing shows that the recombinant proteins are able to withstand room temperature for a week without losing activity. In addition the recombinant proteins were also found to withstand four cycles of freeze and thaw without losing activity.
- How do you convert activity as an ED50 in ng/ml to a specific activity in Units/mg?
- Use formula Specific activity (Units/mg) = 10e6/ ED50 (ng/mL)
- How does the activity of your recombinant proteins compare to competitors?
We quality control each and every lot of recombinant protein. Not only do we check its bioactivity, but we also compare it against other commercially available recombinant proteins. We make sure each recombinant protein’s activity is at least as good as or better than the competition’s. In order to provide you with the best possible product, we ensure that our testing process is rigorous and thorough. If you’re curious and eager to make the switch to BioLegend recombinants, contact your sales representative today!
- What is the specific activity or ED50 of my recombinant protein?
The specific activity range of the protein is indicated on the product datasheets. Because the exact activity values on a per unit basis can largely fluctuate depending on a number of factors, including the nature of the assay, cell density, age of cells/passage number, culture media used, and end user technique, the specific activity is best defined as a range and we guarantee the specific activity of all our lots will be within the range indicated on the datasheet. Please note this only applies to recombinants labeled for use in bioassays. ELISA standard recombinant proteins are not recommended for bioassay usage as they are not tested for these applications.