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BUV563 Streptavidin
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This product is the replacement for [565765].
BUV563 Streptavidin

Flow Cytometric Analysis of Mouse CD45R/B220 or Human CD3 expression using BD Horizon™ BUV563 Streptavidin

Left Panel - Flow cytometric analysis of CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were stained with either Biotin Rat IgG2a, κ Isotype Control (Cat. No. 555842; dashed line histogram) or Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553085/553086; solid line histogram).  The cells were then washed and secondarily stained with BD Horizon BUV563 Streptavidin (Cat. No.  565765). The fluorescence histogram showing CD45R/B220 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes.

Right Panel - Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with either Biotin Mouse IgG1 κ Isotype Control (Cat. No. 555747; dashed line histogram) or Biotin Mouse Anti-Human CD3 antibody (Cat. No. 555331; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The cells were then washed and secondarily stained with BD Horizon BUV563 Streptavidin.   The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System.

Flow Cytometric Analysis of Mouse CD45R/B220 or Human CD3 expression using BD Horizon™ BUV563 Streptavidin

Left Panel - Flow cytometric analysis of CD45R/B220 expression on mouse splenocytes. Mouse splenic leucocytes were stained with either Biotin Rat IgG2a, κ Isotype Control (Cat. No. 555842; dashed line histogram) or Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553085/553086; solid line histogram).  The cells were then washed and secondarily stained with BD Horizon BUV563 Streptavidin (Cat. No.  565765). The fluorescence histogram showing CD45R/B220 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes.

Right Panel - Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with either Biotin Mouse IgG1 κ Isotype Control (Cat. No. 555747; dashed line histogram) or Biotin Mouse Anti-Human CD3 antibody (Cat. No. 555331; solid line histogram). Erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The cells were then washed and secondarily stained with BD Horizon BUV563 Streptavidin.   The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System.

製品詳細
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BD Horizon™
Flow cytometry (Routinely Tested)
0.1 mg/ml
AB_2869708
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


調製と保管

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. Streptavidin was conjugated with BD Horizon BUV563 under optimum conditions, and unconjugated streptavidin and free BD Horizon BUV563 were removed.

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 CompBeads. This will 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).

製品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. CF™ is a trademark of Biotium, Inc.
  5. 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.
  6. BD Horizon Brilliant Ultraviolet 563 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.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
抗体の詳細
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Streptavidin is a non-glycosylated protein that is prepared chromatographically from the bacterium Streptomyces avidinii. Streptavidin homotetramers have a particularly high, non-covalent binding affinity for biotin. When conjugated with fluorochromes, streptavidin has been widely used with biotin-conjugated antibodies and other biotinylated specific-binding molecules (eg, recombinant proteins and lectins) to stain cells and tissues for subsequent multiparameter analysis by flow cytometry, fluorescence microscopy and imaging. Likewise, when conjugated with an enzyme (eg, Horseradish Peroxidase or Alkaline Phosphatase) and coupled with a colorimetric or luminescent substrate development system, streptavidin has found widespread use along with biotinylated antibodies in a number of applications including Western blot, ELISA, ELISPOT, immunocytochemistry and immunohistochemistry.

Streptavidin was conjugated to BD Horizon BUV563 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 which has an Ex Max of 348 nm and an acceptor dye. The tandem has an Em Max at 563 nm. BD Horizon BUV563 can be excited by the 355 nm ultraviolet laser. On instruments with a 561 nm Yellow-Green laser, the recommended bandpass filter is 585/15 nm with a 535 nm long pass to minimize laser light leakage. When BD Horizon BUV563 is used with an instrument that does not have a 561 nm laser, a 560/40 nm filter with a 535 nm long pass may be more optimal. Due to the excitation and emission characteristics of the acceptor dye, there may be spillover into the PE and PE-CF594 detectors. However, the spillover can be corrected through compensation as with any other dye combination.

フォーマットの詳細
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BUV563
The BD Horizon Brilliant™ Ultraviolet 563 (BUV563) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 564-nm. BUV563, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 560-nm (e.g., a 560/40 or a 585/15-nm bandpass filter). The acceptor dye can be excited by the Blue (488-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV563
Ultraviolet 355 nm
350 nm
564 nm
引用&参考文献
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開発者向け参考資料 (1)

  1. Diamandis EP, Christopoulos TK. The biotin-(strept)avidin system: principles and applications in biotechnology. Clin Chem. 1991; 37(5):625-636. (Methodology). 参考文献を見る

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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.