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      V450 Mouse anti-BrdU
      V450 Mouse anti-BrdU
      Flow cytometric analysis of DNA synthesis by TK-1 cells using BD Horizon™ V450 anti-BrdU antibody (Cat. No. 560810). 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 BD Horizon™ V450 anti-BrdU antibody followed by the DNA-specific dye, 7-AAD (7-AAD Staining Solution, Cat. No. 559925). Two-color flow cytometric dot plots showing the correlated expression patterns of 7-AAD 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™ LSR II System.
      V450 Mouse anti-BrdU
      Flow cytometric analysis of DNA synthesis by TK-1 cells using BD Horizon™ V450 anti-BrdU antibody (Cat. No. 560810). 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 BD Horizon™ V450 anti-BrdU antibody followed by the DNA-specific dye, 7-AAD (7-AAD Staining Solution, Cat. No. 559925). Two-color flow cytometric dot plots showing the correlated expression patterns of 7-AAD 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™ LSR II System.
      Product Details
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      BD Horizon™
      5-bromo-2'-deoxyuridine, 5-Bromouracil deoxyriboside, BUdR
      Mouse (QC Testing), Human (Tested in Development)
      Mouse IgG1, κ
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      AB_2033930
      Aqueous buffered solution containing protein stabilizer 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 to the dye under optimum conditions and unreacted dye was removed.
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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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. Pacific Blue™ is a trademark of Life Technologies Corporation.
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      560810 Rev. 2
      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.

      The antibody is conjugated to BD HorizonTM V450, which has been developed for use in multicolor flow cytometry experiments and is available exclusively from BD Biosciences. It is excited by the Violet laser Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD HorizonTM V450 can be used in place of Pacific Blue™ conjugates.

      560810 Rev. 2
      Format Details
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      V450
      BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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      V450
      Violet 405 nm
      405 nm
      450 nm
      560810 Rev.2
      Citations & References
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      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).
      560810 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.

      Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.

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