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 (including BD OptiBuild Brilliant reagents) 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 antibody was developed for use in flow cytometry.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- BD Horizon Brilliant Violet 650 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- Alexa Fluor® is a registered trademark of Life Technologies Corporation.
Companion Products
The 1A5 monoclonal antibody specifically recognizes Sialic acid binding Ig-like lectin 5 (Siglec-5) which is also known as CD170, CD33 antigen-like 2 (CD33L2) and Obesity binding protein-2 (OBBP2). CD170 is a type I transmembrane glycoprotein that forms a ~140 kDa dimer on the cell surface. CD170 is encoded by SIGLEC5 which belongs to the SIGLEC (sialic acid binding Ig-like lectin) family within the immunoglobulin superfamily. The extracellular region of CD170 consists of an N-terminal V-set Ig-like domain, which contains the sialic acid binding site, followed by 3 C2-set Ig like domains, a transmembrane region and a cytoplasmic tail containing 2 immunoreceptor tyrosine-based inhibitory motifs (ITIMs) which regulate cellular signaling. CD170 bind to alpha-2,3- and alpha-2,6-linked sialic acid glycoproteins and glycolipids and to glycophorin A. It is highly expressed on neutrophils and expressed at lower levels on monocytes, macrophages, dendritic cells, and some lymphocytes. CD170 is an adhesion molecule that may function in regulating cellular activation and in sialic-acid dependent binding to cells.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon Brilliant™ Violet family of dyes. This dye is a tandem fluorochrome of BD Horizon BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
Development References (5)
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Arase N, Takeuchi A, Unno M, et al. Heterotypic interaction of CRTAM with Necl2 induces cell adhesion on activated NK cells and CD8+ T cells.. Int Immunol. 2005; 17(9):1227-37. (Immunogen: Flow cytometry). View Reference
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Fuchs A, Colonna M. The role of NK cell recognition of nectin and nectin-like proteins in tumor immunosurveillance.. Semin Cancer Biol. 2006; 16(5):359-66. (Biology). View Reference
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Kennedy J, Vicari AP, Saylor V, et al. A molecular analysis of NKT cells: identification of a class-I restricted T cell-associated molecule (CRTAM).. J Leukoc Biol. 2000; 67(5):725-34. (Biology). View Reference
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Takeuchi A, Badr Mel S, Miyauchi K, et al. CRTAM determines the CD4+ cytotoxic T lymphocyte lineage.. J Exp Med. 2016; 213(1):123-38. (Clone-specific: Flow cytometry). View Reference
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Takeuchi A, Itoh Y, Takumi A, et al. CRTAM confers late-stage activation of CD8+ T cells to regulate retention within lymph node.. J Immunol. 2009; 183(7):4220-8. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
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.