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      Alexa Fluor® 647 Mouse Anti-Human BMI-1
      Alexa Fluor® 647 Mouse Anti-Human BMI-1
      Flow cytometric analysis of BMI-1 expression in a human osteosarcoma cell line. U-2 OS cells  (ATCC; HTB-96™) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human BMI-1 antibody (Cat. No. 562637; solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
      Alexa Fluor® 647 Mouse Anti-Human BMI-1
      Image analysis of BMI-1 expressed in human osteosarcoma cell line. U-2 OS cells (ATCC, HTB-96™) were fixed with BD Cytofix™ Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma, Cat. No. X-100), and stained with Alexa Fluor® 647 Mouse Anti-Human-BMI-1 antibody (Cat. No. 562637; 5 μg/ml; pseudo-colored red). Counter-staining was with DAPI (pseudo-colored blue). The image was captured using a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
      Alexa Fluor® 647 Mouse Anti-Human BMI-1 Alexa Fluor® 647 Mouse Anti-Human BMI-1
      Flow cytometric analysis of BMI-1 expression in a human osteosarcoma cell line. U-2 OS cells  (ATCC; HTB-96™) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655) and permeabilized with BD Phosflow™ Perm Buffer III (Cat. No. 558050). The cells were stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557783; dashed line histogram) or Alexa Fluor® 647 Mouse Anti-Human BMI-1 antibody (Cat. No. 562637; solid line histogram) at matched concentrations. The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of intact cells. Flow cytometry was performed on a BD™ LSR II Flow Cytometry System.
      Image analysis of BMI-1 expressed in human osteosarcoma cell line. U-2 OS cells (ATCC, HTB-96™) were fixed with BD Cytofix™ Buffer (Cat. No. 554655), permeabilized with 0.1% Triton™ X-100 (Sigma, Cat. No. X-100), and stained with Alexa Fluor® 647 Mouse Anti-Human-BMI-1 antibody (Cat. No. 562637; 5 μg/ml; pseudo-colored red). Counter-staining was with DAPI (pseudo-colored blue). The image was captured using a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ Software.
      Product Details
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      BD Pharmingen™
      BMI1; PCGF4; polycomb group ring finger 4; RING finger protein 51; RNF51
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Human BMI-1 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence (Tested During Development)
      5 µl
      648
      AB_2737693
      Aqueous buffered solution containing BSA, 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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
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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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      5. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      8. 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.
      9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      10. Triton is a trademark of the Dow Chemical Company.
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      562637 Rev. 1
      Antibody Details
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      P51-311

      The P51-311 monoclonal antibody binds to human BMI-1 (B lymphoma Mo-MLV insertion region 1 homolog). BMI1 is a c-myc cooperating oncogene that encodes an ~45 kDa protein that is a member of the Polycomb Group (PcG) of proteins. PcG proteins are essential for the maintenance, but not initiation, of the transcriptionally repressed state of certain developmental genes. PcG proteins are a structurally diverse group of proteins with conserved functions from fly to human cells. PcG proteins form complexes and regulate the expression of genes involved in cell cycle, DNA repair and differentiation that are crucial for maintaining the self renewal of normal and cancer stem cells. Specifically, BMI-1 is a core component of PRC1 (polycomb repressive complex 1). BMI-1, via the up-regulation of hTERT and independent of c-myc, can immortalize mammary epithelial cells. BMI-1 has also been shown to repress the INK4A locus that controls the tumor suppressors p16 and p19ARF (mouse homologue of p14ARF) in mouse models. BMI-1 plays a role in maintaining the self-renewal capacities of stem cells including hematopoietic, intestinal, retinal and neural stem cells.  During antibody development, the purified P51-311 monoclonal antibody was found to detect BMI-1 by Western blot analysis of cellular lysates and by indirect immunofluorescent staining and flow cytometric analysis of fixed and permeabilized cells.

      562637 Rev. 1
      Format Details
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      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      562637 Rev.1
      Citations & References
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      Development References (6)

      1. Dimri GP, Martinez JL, Jacobs JJ. The Bmi-1 oncogene induces telomerase activity and immortalizes human mammary epithelial cells. Cancer Res. 2002; 62(16):4736-4745. (Biology). View Reference
      2. Itahana K, Zou Y, Itahana Y. Control of the replicative life span of human fibroblasts by p16 and the polycomb protein Bmi-1. Mol Cell Biol. 2003; 23(1):389-401. (Biology). View Reference
      3. Molofsky AV, Pardal R, Iwashita T, Park IK, Clarke MF, Morrison SJ. Bmi-1 dependence distinguishes neural stem cell self-renewal from progenitor proliferation. Nature. 2003; 6961(962):967. (Biology). View Reference
      4. Park IK, Qian D, Kiel M, et al. Bmi-1 is required for maintenance of adult self-renewing haematopoietic stem cells. Nature. 2003; 423(6937):302-305. (Biology). View Reference
      5. Sangiorgi, E., Capecchi, M. R. Bmi1 is expressed in vivo in intestinal stem cells. Nat Genet. 2008; 40(7):915-920. (Biology). View Reference
      6. Tian H, Biehs B, Warming S, et al. A reserve stem cell population in small intestine renders Lgr5-positive cells dispensable. Nature. 2011; 478(7368):255-259. (Biology). View Reference
      View All (6) View Less
      562637 Rev. 1

      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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