Preparation and Storage
推奨アッセイ手順
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 CompBead 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 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. 563794/566349) 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).
- BD Horizon Brilliant Ultraviolet 661 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.
関連製品
The p282 (HI9) monoclonal antibody specifically binds to CD59, a 19 kDa glycosylphosphatidylinositol (GPI)-anchored glycoprotein, expressed on hematopoietic and non-hematopoietic cells. Because of its interaction with complement activated products, CD59 has been termed membrane-attack-complex-inhibitory factor (MACIF), homologus restriction factor (HRF20), membrane inhibitor of reactive lysis (MIRL) and Protectin. It inhibits the cytolytic activity of the complement system by binding to C8 and C9, thereby blocking the assembly of the membrane attack complex. CD59 also participates in spontaneous T-cell/erythrocyte adhesion, interacts with CD2, and plays a role in T-cell activation.
The antibody was conjugated to BD Horizon BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP. Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).
Development References (8)
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Davies A, Lachmann PJ. Membrane defence against complement lysis: the structure and biological properties of CD59. Immunol Res. 1993; 12(3):258-275. (Biology). View Reference
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Deckert M, Kubar J, Bernard A. CD58 and CD59 molecules exhibit potentializing effects in T cell adhesion and activation. J Immunol. 1992; 148(3):672-677. (Clone-specific: Blocking, Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
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Deckert M, Kubar J, Zoccola D, et al. CD59 molecule: a second ligand for CD2 in T cell adhesion. Eur J Immunol. 1992; 22(11):2943-2947. (Clone-specific: Blocking). View Reference
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Groux H, Huet S, Aubrit F, Tran HC, Boumsell L, Bernard A. A 19-kDa human erythrocyte molecule H19 is involved in rosettes, present on nucleated cells, and required for T cell activation. Comparison of the roles of H19 and LFA-3 molecules in T cell activation. J Immunol. 1989; 142(9):3013-3020. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Inhibition). View Reference
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
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Whitlow MB, Iida K, Stefanova I, Bernard A, Nussenzweig V. H19, a surface membrane molecule involved in T-cell activation, inhibits channel formation by human complement. Cell Immunol. 1990; 126(1):176-184. (Biology). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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.