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      PerCP-Cy™5.5 Mouse anti-BrdU
      PerCP-Cy™5.5 Mouse anti-BrdU
      Flow cytometric analysis of DNA synthesis by TK-1 cells using PerCP-Cy5.5 anti-BrdU antibody. TK-1 cells were either pulsed with 50 µM BrdU for 1 hour (left panel) or were not pulsed (right panel). Staining was performed using the procedure from the BD Pharmingen™ FITC BrdU Flow Kit (Cat. No. 559619). The permeabilized cells were stained with the PerCP-Cy5.5 anti-BrdU antibody followed by the DNA-specific dye, DAPI dihydrochloride at 1 µg/ml (Sigma, Cat. No. D9542). Two-color flow cytometric dot plots showing the correlated expression patterns of DAPI vs BrdU were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed with doublet discrimination using a BD™ LSRII System.
      PerCP-Cy™5.5 Mouse anti-BrdU
      Flow cytometric analysis of DNA synthesis by TK-1 cells using PerCP-Cy5.5 anti-BrdU antibody. TK-1 cells were either pulsed with 50 µM BrdU for 1 hour (left panel) or were not pulsed (right panel). Staining was performed using the procedure from the BD Pharmingen™ FITC BrdU Flow Kit (Cat. No. 559619). The permeabilized cells were stained with the PerCP-Cy5.5 anti-BrdU antibody followed by the DNA-specific dye, DAPI dihydrochloride at 1 µg/ml (Sigma, Cat. No. D9542). Two-color flow cytometric dot plots showing the correlated expression patterns of DAPI vs BrdU were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometry was performed with doublet discrimination using a BD™ LSRII System.
      Product Details
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      BD Pharmingen™
      5-bromo-2'-deoxyuridine, 5-Bromouracil deoxyriboside, BUdR
      Mouse (QC Testing), Human,Rat (Tested in Development)
      Mouse IgG1, κ
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2033929
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      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. The antibody was conjugated with PerCP-Cy5.5 under optimum conditions, and unconjugated antibody and free PerCP-Cy5.5 were removed. Storage of PerCP-Cy5.5 conjugates in unoptimized diluent is not recommended and may result in loss of signal intensity.
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      Recommended Assay Procedures

      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 to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
      3. PerCP-Cy5.5 is optimized for use with a single argon ion laser emitting 488-nm light. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using dual-laser cytometers, which may directly excite both PerCP and Cy5.5™. We recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
      4. PerCP-Cy5.5–labelled antibodies can be used with FITC- and R-PE–labelled reagents in single-laser flow cytometers with no significant spectral overlap of PerCP-Cy5.5, FITC, and R-PE fluorescence.
      5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. 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.
      8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
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      560809 Rev. 3
      Antibody Details
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      3D4

      Bromodeoxyuridine (BrdU) is an analog of thymidine that can be incorporated into newly synthesized DNA by cells entering and progressing through the DNA synthesis (S) phase of the cell cycle.  The amount of incorporated BrdU depends on the amount of time that the cells are exposed to BrdU (pulse time), the rate of cell division, and whether the cells are in early, mid, or late S phase. Investigators can identify cycling cells in an asynchronous cell population and determine cell cycle kinetics by detecting incorporated BrdU.

      The 3D4 monoclonal antibody reacts with BrdU, but not other nucleotides, in single-stranded DNA. Random cleavage (nicking) of cellular DNA with DNase I permits the binding of the antibody to incorporated BrdU.

      560809 Rev. 3
      Format Details
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      PerCP-Cy5.5
      PerCP-Cy5.5 dye is part of the BD blue family of dyes. This tandem fluorochrome is comprised of a fluorescent protein complex (PerCP) with an excitation maximum (Ex Max) of 482 nm and an acceptor dye with an emission maximum (Em Max) at 676 nm. PerCP-Cy5 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 680 nm (e.g., a 695/40 nm bandpass filter). The donor dye can be partially excited by the Violet (405-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.
      altImg
      PerCP-Cy5.5
      Blue 488 nm
      482 nm
      676 nm
      560809 Rev.3
      Citations & References
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      View product citations for antibody "560809" on CiteAb

      Development References (3)

      1. Dolbeare F, Gratzner H, Pallavicini MG, Gray JW. Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc Natl Acad Sci U S A. 1983; 80(18):5573-5577. (Methodology: Flow cytometry). View Reference
      2. Keren DF, Hanson CA, Hurtubise PE. David F. Keren, Curtis A. Hanson, Paul E. Hurtubise., ed. Flow cytometry and clinical diagnosis. Chicago: ASCP Press; 1994:1-676.
      3. Miltenburger HG, Sachse G, Schliermann M. S-phase cell detection with a monoclonal antibody. Dev Biol Stand. 1987; 66:91-99. (Clone-specific: Immunofluorescence).
      560809 Rev. 3

       

      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.