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BB515 Streptavidin
BB515 Streptavidin

Flow cytometric analysis of CD45R/B220 expression on mouse lymphocytes using Biotin Rat Anti-Mouse CD45R/B220 and BD Horizon™ BB515 Streptavidin.   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 either not stained with a primary antibody (dashed line histogram) or stained with Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553085/553086). The cells were washed and stained with BD Horizon BB515 Streptavidin (Cat. No. 564453). Flow cytometric histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSRII System.

Flow cytometric analysis of CD45R/B220 expression on mouse lymphocytes using Biotin Rat Anti-Mouse CD45R/B220 and BD Horizon™ BB515 Streptavidin.   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 either not stained with a primary antibody (dashed line histogram) or stained with Biotin Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553085/553086). The cells were washed and stained with BD Horizon BB515 Streptavidin (Cat. No. 564453). Flow cytometric histograms were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD™ LSRII System.

製品詳細
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BD Horizon™
Flow cytometry (Routinely Tested)
0.1 mg/ml
AB_2869580
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 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 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.

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).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
  8. 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 (BSA) RUO
サイズ 500 mL カタログ番号 554657
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Stain Buffer (FBS) RUO
サイズ 500 mL カタログ番号 554656
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Lysing Buffer RUO
サイズ 100 mL カタログ番号 555899
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Biotin Rat Anti-Mouse CD45R/B220 RUO
サイズ 0.1 mg カタログ番号 553085
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564453 Rev. 3
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 BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

564453 Rev. 3
フォーマットの詳細
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BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BB515
Blue 488 nm
490 nm
515 nm
564453 Rev.3
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.
564453 Rev. 3

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