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BV480 Streptavidin
BV480 Streptavidin

Immunofluorescence staining of human U-2 OS cells. Cultured cells from the human U-2 OS (Osteosarcoma, ATCC HTB-96) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized (5 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050), washed with 1x PBS, and blocked with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1x PBS. The cells were then treated using the Avidin/Biotin Blocking Kit (Vector Laboratories), and stained with Biotin Anti-PCNA antibody (Abcam). After washing, the cells were stained with the second step reagent, BD Horizon™ BV480 Streptavidin (Cat. No. 564876) (pseudo-colored green) at 2.5 μg/mL. Cells were then stained with Alexa Fluor® 488 Mouse Anti-β-Tubulin antibody (Cat No. 558605) (pseudo-colored red). DAPI (Cat. No. 564907) was used as a nuclear counterstain (pseudo-colored blue). Slides were mounted with ProLong® Gold and the image was captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software. 20X objective.

Immunofluorescence staining of human U-2 OS cells. Cultured cells from the human U-2 OS (Osteosarcoma, ATCC HTB-96) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized (5 min) with BD Phosflow™ Perm Buffer III (Cat. No. 558050), washed with 1x PBS, and blocked with 5% Goat serum, 1% BSA, and 0.5% Triton™ X-100 diluted in 1x PBS. The cells were then treated using the Avidin/Biotin Blocking Kit (Vector Laboratories), and stained with Biotin Anti-PCNA antibody (Abcam). After washing, the cells were stained with the second step reagent, BD Horizon™ BV480 Streptavidin (Cat. No. 564876) (pseudo-colored green) at 2.5 μg/mL. Cells were then stained with Alexa Fluor® 488 Mouse Anti-β-Tubulin antibody (Cat No. 558605) (pseudo-colored red). DAPI (Cat. No. 564907) was used as a nuclear counterstain (pseudo-colored blue). Slides were mounted with ProLong® Gold and the image was captured on a BD Pathway™ 435 Cell Analyzer (epifluorescence microscope) and merged using BD Attovision™ Software. 20X objective.

製品詳細
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BD Horizon™
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.1 mg/ml
AB_2869619
Aqueous buffered solution containing BSA 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 BD Horizon BV480 under optimum conditions, and unconjugated streptavidin and free BD Horizon BV480 were removed.

推奨アッセイ手順

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).

For Immunofluorescence Applications:

The use of a mounting reagent (eg, ProLong® Gold) is highly recommended to maximize the photostability of BV480.  For confocal microscopy systems, a 440 nm laser is the optimal excitation source and the recommended emission filter is a 485/20 nm bandpass filter.  

For epifluorescence microscopes with broad spectrum excitation sources,  the recommended excitation and emission filters are 445/20 nm and 485/20 nm bandpass filters, respectively.  For specific multicolor imaging applications, the exact filter configurations should be optimized by the end user. For additional instrument/filter configuration information, please visit http://www.bdbiosciences.com/research/cellularimaging.

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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. Triton is a trademark of the Dow Chemical Company.
  5. ProLong® is a registered trademark of Thermo Fisher Scientific, Inc. Waltham, MA.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564876 Rev. 4
抗体の詳細
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BD Horizon BV480 Streptavidin is a useful detection reagent for labeling biotinylated primary or secondary antibodies for detection of antigens by immunofluorescence. Streptavidin has very high binding affinity for biotin, which is conjugated to the antibody.

Streptavidin was conjugated to BD Horizon BV480 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 436-nm and Em Max at 478-nm, BD Horizon BV480 can be excited by the violet laser and detected in the BD Horizon BV510 (525/40-nm) filter set.  BV480 has less spillover into the BV605 detector and, in general, is brighter than BV510.

564876 Rev. 4
フォーマットの詳細
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BV480
Violet 405 nm
440 nm
479 nm
564876 Rev.4
引用&参考文献
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開発者向け参考資料 (10)

  1. Afar B, Merrill J, Clark EA. Detection of lymphocyte subsets using three-color/single-laser flow cytometry and the fluorescent dye peridinin chlorophyll-alpha protein. J Clin Immunol. 1991; 11(5):254-261. (Biology). 参考文献を見る
  2. Beavis AJ, Pennline KJ. Allo-7: a new fluorescent tandem dye for use in flow cytometry. Cytometry. 1996; 24(4):390-395. (Biology). 参考文献を見る
  3. 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
  4. Greimers R, Trebak M, Moutschen M, Jacobs N, Boniver J. Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets. Cytometry. 1996; 23(3):205-217. (Biology). 参考文献を見る
  5. Roederer M, Kantor AB, Parks DR, Herzenberg LA. Cy7PE and Cy7APC: bright new probes for immunofluorescence. Cytometry. 1996; 24(3):191-197. (Biology). 参考文献を見る
  6. Shapiro HM. Practical flow cytometry, 4th ed.. Hoboken, N.J.: Wiley-Liss; 2003:1-681.
  7. Stewart CC, Stewart SJ. Immunological monitoring utilizing novel probes. Ann N Y Acad Sci. 1993; 677:94-112. (Biology).
  8. Takizawa F, Kinet JP, Adamczewski M. Binding of phycoerythrin and its conjugates to murine low affinity receptors for immunoglobulin G. J Immunol Methods. 1993; 162(2):269-272. (Biology). 参考文献を見る
  9. Waggoner AS, Ernst LA, Chen CH, Rechtenwald DJ. PE-CY5. A new fluorescent antibody label for three-color flow cytometry with a single laser. Ann N Y Acad Sci. 1993; 677:185-193. (Biology). 参考文献を見る
  10. van Vugt MJ, van den Herik-Oudijk IE, van de Winkle JG. Binding of PE-CY5 conjugates to the human high-affinity receptor for IgG (CD64). Blood. 1996; 88(6):2358-2361. (Biology). 参考文献を見る
すべて表示する (10) 表示項目を減らす
564876 Rev. 4

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.

23-22944-00