Preparation And Storage
Recommended Assay Procedures
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
The 2-69 monoclonal antibody specifically binds to 2B4 which is also known as CD244. CD244 is an approximately 63-70 kDa type I transmembrane glycoprotein. It is a SLAM submember (SLAMF4) and CD2 family member of the immunoglobulin superfamily. CD244 was originally identified in the mouse as a non-MHC-restricted cytotoxicity mediator present on NK cells and CD8+ T cells. In humans, CD244 is expressed on NK cells, γδ T cells, subsets of effector and memory CD8+ T cells, monocytes, eosinophils and basophils. This expression pattern suggests a broad role for CD244 in the regulation of leukocyte activation. CD244 binds CD48 with high affinity. CD244 may serve both stimulatory and inhibitory functions depending on a number of factors. These include the intensity and duration of CD244 interactions with its ligands as well as the presence of other signals generated through different receptor-ligand interactions. 2B4 was clustered as CD244 in the VIIth HLDA workshop.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (7)
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Boles KS, Nakajima H, Colonna M, et al. Molecular characterization of a novel human natural killer cell receptor homologous to mouse 2B4. Tissue Antigens. 1999; 54(1):27-34. (Biology). View Reference
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Brown MH, Boles K, van der Merwe PA, Kumar V, Mathew PA, Barclay AN. 2B4, the natural killer and T cell immunoglobulin superfamily surface protein, is a ligand for CD48. J Exp Med. 1998; 188(11):2083-2090. (Biology). View Reference
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Colonna M, Nakajima H, Cella M. Inhibitory and activating receptors involved in immune surveillance by human NK and myeloid cells. J Leukoc Biol. 1999; 66(5):718-722. (Biology). View Reference
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Latchman Y, McKay PF, Reiser H. Identification of the 2B4 molecule as a counter-receptor for CD48. J Immunol. 1998; 161(11):5809-5812. (Biology). View Reference
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Nakajima H, Cella M, Bouchon A, et al. Patients with X-linked lymphoproliferative disease have a defect in 2B4 receptor-mediated NK cell cytotoxicity. Eur J Immunol. 2000; 30(11):3309-3318. (Immunogen: Flow cytometry, Functional assay, Immunoprecipitation, Stimulation, Western blot). View Reference
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Nakajima H, Cella M, Langen H, Friedlein A, Colonna M. Activating interactions in human NK cell recognition: the role of 2B4-CD48. Eur J Immunol. 1999; 29(5):1676-1683. (Biology). View Reference
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Tassi I, Colonna M. The cytotoxicity receptor CRACC (CS-1) recruits EAT-2 and activates the PI3K and phospholipase Cgamma signaling pathways in human NK cells. J Immunol. 2005; 175(12):7996-8002. (Clone-specific: Immunoprecipitation). View Reference
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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.