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APC-R700 Streptavidin
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APC-R700 Streptavidin

Flow cytometric analysis of mouse CD45R/B220 or human CD3 expression on leucocytes using Biotin-conjugated primary antibodies and BD Horizon™ APC-R700 Streptavidin.

Left Plot - Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then 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 washed and stained with BD Horizon APC-R700 Streptavidin (Cat. No. 565144). 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 leucocytes.

Right Plot - 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 washed and stained with BD Horizon APC-R700 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 on leucocytes using Biotin-conjugated primary antibodies and BD Horizon™ APC-R700 Streptavidin.

Left Plot - Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then 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 washed and stained with BD Horizon APC-R700 Streptavidin (Cat. No. 565144). 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 leucocytes.

Right Plot - 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 washed and stained with BD Horizon APC-R700 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_2869657
Aqueous buffered solution containing protein stabilizer and ≤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 dye under optimum conditions, and unconjugated Streptavidin and free dye were removed

推奨アッセイ手順

BD Horizon™ APC-R700 Streptavidin is a useful second-step reagent for the indirect immunofluorescent staining of cells in combination with biotinylated primary antibodies for flow cytometric analysis.

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

コンパニオン製品

Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.1 mg カタログ番号 553141
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Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
サイズ 0.5 mg カタログ番号 553142
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Stain Buffer (FBS) RUO
サイズ 500 mL カタログ番号 554656
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Stain Buffer (BSA) RUO
サイズ 500 mL カタログ番号 554657
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Biotin Rat IgG2a, κ Isotype Control RUO
サイズ 100 Tests カタログ番号 555842
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Biotin Rat Anti-Mouse CD45R/B220 RUO
サイズ 0.1 mg カタログ番号 553085
sampleImage/
565144 Rev. 2
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 APC-R700, which has been developed exclusively by BD Biosciences as a better alternative to Alexa Fluor® 700. APC-R700 excites and emits at similar wavelengths to Alexa Fluor® 700 yet exhibits significantly improved brightness. This dye can be excited by the red laser and detected with the same filter set as Alexa Fluor® (eg, 730/45-nm filter).

565144 Rev. 2
フォーマットの詳細
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APC-R700
The BD Horizon™ APC-R700 (APC-R700) Dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of an Allophycocyanin (APC) dye donor that has excitation maximum (Ex Max) of 651-nm and an acceptor dye, R700, with an emission maximum (Em Max) at 706-nm. APC-R700, driven by BD innovation, is designed to be excited by the red (627–640-nm) laser and detected using an optical filter centered near 710-nm (e.g., a 720/40-nm bandpass filter). APC-R700 is a brighter alternative to Alexa Fluor™ 700. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC-R700
Red 627-640 nm
651 nm
706 nm
565144 Rev.2
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
565144 Rev. 2

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.