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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 PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse anti-Human BMI-1 antibody (Cat. No. 562650; 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 using a BD™ LSR II Flow Cytometry System.
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 PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse anti-Human BMI-1 antibody (Cat. No. 562650; 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 using a BD™ LSR II Flow Cytometry System.
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 PerCP-Cy™5.5 Mouse IgG1, κ Isotype Control (Cat. No. 550795; dashed line histogram) or PerCP-Cy™5.5 Mouse anti-Human BMI-1 antibody (Cat. No. 562650; 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 using a BD™ LSR II Flow Cytometry System.
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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.
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