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      PE Mouse Anti- BrdU Set

      BD Pharmingen™ PE Mouse Anti- BrdU Set

      (RUO)
      PE Mouse Anti- BrdU Set
      Two-color flow cytometric analysis of DNA synthesis by REH cells. REH  (Human leukemia, ATCC CRL-8286) cells were labeled with  1mM BrdU for one hour, washed, fixed with 75% ice cold ethanol and permeabilized with Tween-20.  The  cells were then stained with either PE Mouse IgG1 κ Isotype Control  (Left Panel) or PE Mouse Anti-BrdU antibody (Right Panel; Cat No. 556029) followed by staining with  DAPI Solution (Cat. No. 564907; nucleic acid stain).  Two-color flow cytometric dot plots showing the correlated expression patterns of DAPI (total cellular DNA) versus incorporated BrdU (representing newly-synthesized DNA) were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II System
      PE Mouse Anti- BrdU Set PE Mouse Anti- BrdU Set
      Two-color flow cytometric analysis of DNA synthesis by REH cells. REH  (Human leukemia, ATCC CRL-8286) cells were labeled with  1mM BrdU for one hour, washed, fixed with 75% ice cold ethanol and permeabilized with Tween-20.  The  cells were then stained with either PE Mouse IgG1 κ Isotype Control  (Left Panel) or PE Mouse Anti-BrdU antibody (Right Panel; Cat No. 556029) followed by staining with  DAPI Solution (Cat. No. 564907; nucleic acid stain).  Two-color flow cytometric dot plots showing the correlated expression patterns of DAPI (total cellular DNA) versus incorporated BrdU (representing newly-synthesized DNA) were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD™ LSR II System
      Product Details
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      BD Pharmingen™
      Human (QC Testing), Mouse,Rat (Tested in Development)
      Flow cytometry, Intracellular staining (flow cytometry) (Routinely Tested)
      RUO
      AB_396305


      Description

      Bromodeoxyuridine (BrdU) is an analog of thymidine that can be incorporated into newly synthesized DNA by cells entering and progressing thru the DNA synthesis (S) phase of the cell cycle.  The amount of BrdU that gets incorporated is dependent upon 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.  Detection of incorporated BrdU allows the investigator to identify cycling cells in an asynchronous cell population and to determine cell cycle kinetics.

      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.

      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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
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      Recommended Assay Procedures

      Investigators:  Please note that this product is routinely tested on fixed and permeabilized REH cell line with 70-80% cold ethanol. Please see the detailed protocol example at http://www.bdbiosciences.com/us/resources/protocols/s/brdudetection  This product is NOT recommended to use with the BrdU flow kits (e.g. Cat. Nos. 559619, 552598, 557892 and 557892).

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
      7. An isotype control should be used at the same concentration as the antibody of interest.
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      556029 Rev. 10
      Components
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      Description Quantity/Size Part Number EntrezGene ID
      PE Mouse IgG1 κ Isotype Control 100 Tests (1 ea) 51-35405X-2 N/A
      PE Mouse Anti-BrdU 100 Tests (1 ea) 51-33285X N/A
      556029 Rev. 10
      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. (Biology). 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).
      556029 Rev. 10

      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.

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