Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Companion Products
The 6F7 monoclonal antibody specifically recognizes CD272 which is also known as B- and T-lymphocyte attenuator (BTLA) and is exclusively expressed on lymphoid cells. CD272 (BTLA) is a type 1 transmembrane glycoprotein that is encoded by Btla (B and T lymphocyte associated). CD272 (BTLA) contains a V-type Ig-like domain in its extracellular region followed by a transmembrane sequence, and a cytoplasmic domain with three tyrosine-based motifs, two immunoreceptor tyrosine-based inhibitory motifs (ITIM) and a Grb-2 recognition consensus sequence. The existence of three distinct BTLA alleles has been reported which encode molecules with different Ig domain structure and expression patterns on lymphoid cell subsets amongst different mouse strains. For example, whereas C57BL/6 and BALB/c mice both variably express CD272 (BTLA) on developing and mature T and B lymphocytes and dendritic cells (DC), C57BL/6 mice, but not BALB/c mice, also express CD272 (BTLA) on NK cells and macrophages. CD272 (BTLA) expression is upregulated by activated T cells including Th1, Th2, and anergic T cells. Herpesvirus entry mediator (HVEM), also known as CD270 and LIGHT-R, has been identified as a ligand for CD272 (BTLA). The crosslinking of CD272 (BTLA) by HVEM inhibits T-cell proliferation and cytokine production. CD272 (BTLA) is structurally like other coinhibitory receptors including CD152/CTLA-4 and CD279/PD-1. Although these coinhibitory receptors and their ligands maintain immunological homeostasis and self-tolerance, they may also serve as immune checkpoint molecules that inhibit adaptive immune responses against tumors and chronic infections.
Development References (7)
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Crawford A, Wherry EJ. Editorial: Therapeutic potential of targeting BTLA.. J Leukoc Biol. 2009; 86(1):5-8. (Clone-specific). View Reference
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Han P, Goularte OD, Rufner K, Wilkinson B, Kaye J. An inhibitory Ig superfamily protein expressed by lymphocytes and APCs is also an early marker of thymocyte positive selection. J Immunol. 2004; 172(10):5931-5939. (Biology). View Reference
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Hurchla MA, Sedy JR, Gavrieli M, Drake CG, Murphy TL, Murphy KM. B and T lymphocyte attenuator exhibits structural and expression and is highly induced in anergic CD4+T cells. J Immunol. 2005; 174(6):3377-3385. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
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Nurieva RI, Chung Y, Martinez GJ, et al. Bcl6 mediates the development of T follicular helper cells. Science. 2009; 325(5943):1001-1005. (Biology). View Reference
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Sedy JR, Gavrieli M, Potter KG, et al. B and T lymphocyte attenuator regulates T cell activation through interaction with herpesvirus entry mediator. Nat Immunol. 2005; 6(1):90-98. (Biology). View Reference
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Watanabe N, Gavrieli M, Sedy JR, et al. BTLA is a lymphocyte inhibitory receptor with similarities to CTLA-4 and PD-1. Nat Immunol. 2003; 4(7):670-679. (Biology). View Reference
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Watanabe N, Nakajima H. Coinhibitory molecules in autoimmune diseases. Clin Dev Immunol. 2012; 2012:1-7. (Biology). 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.