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BUV615 Streptavidin
BUV615 Streptavidin

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™ BUV615 Streptavidin (Cat. No. 613013). 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. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

BUV615 Streptavidin

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). The cells were then washed and secondarily stained with BD Horizon™ BUV615 Streptavidin (Cat. No. 613013). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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 intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System X-20 and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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™ BUV615 Streptavidin (Cat. No. 613013). 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. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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). The cells were then washed and secondarily stained with BD Horizon™ BUV615 Streptavidin (Cat. No. 613013). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). 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 intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System X-20 and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

製品詳細
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BD Horizon™
Flow cytometry (Routinely Tested)
0.1 mg/ml
AB_2870281
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. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Streptavidin was conjugated with dye under optimum conditions, and unconjugated Streptavidin and free dye 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 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).

製品通知

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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 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.
  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.
613013 Rev. 1
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.

The Streptavidin was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP.  Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).

613013 Rev. 1
フォーマットの詳細
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BUV615
The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) 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 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers 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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BUV615
Ultraviolet 355 nm
350 nm
615 nm
613013 Rev.1
引用&参考文献
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開発者向け参考資料 (2)

  1. Diamandis EP, Christopoulos TK. The biotin-(strept)avidin system: principles and applications in biotechnology. Clin Chem. 1991; 37(5):625-636. (Biology). 参考文献を見る
  2. Shapiro HM. Practical flow cytometry, 4th ed.. Hoboken, N.J.: Wiley-Liss; 2003:1-681.
613013 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.