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Flow cytometric analysis of human CD123 expression on cells transfected with human IL3RA. Chinese Hamster Ovary (CHO) cells transfected with human IL3RA were stained with either BD Horizon™ BV650 Mouse Anti-Human CD123 antibody (Cat. No. 563405; solid line histogram) or with a BD Horizon™ BV650 Mouse IgG2a, κ Isotype Control (Cat. No. 563417; dashed line histogram). Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of human CD123 expression on cells transfected with human IL3RA. Chinese Hamster Ovary (CHO) cells transfected with human IL3RA were stained with either BD Horizon™ BV650 Mouse Anti-Human CD123 antibody (Cat. No. 563405; solid line histogram) or with a BD Horizon™ BV650 Mouse IgG2a, κ Isotype Control (Cat. No. 563417; dashed line histogram). Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of human CD123 expression on cells transfected with human IL3RA. Chinese Hamster Ovary (CHO) cells transfected with human IL3RA were stained with either BD Horizon™ BV650 Mouse Anti-Human CD123 antibody (Cat. No. 563405; solid line histogram) or with a BD Horizon™ BV650 Mouse IgG2a, κ Isotype Control (Cat. No. 563417; dashed line histogram). Fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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The 7G3 monoclonal antibody specifically recognizes human CD123, the 70 kDa IL-3 Receptor α (IL-3Rα) chain. CD123 associates with CD131, the 120-140 kDa Common β chain to form the IL-3 Receptor Complex. CD131 is shared with the receptors for interleukins IL-5 and GM-CSF. IL-3Rα is expressed on hematopoietic progenitors and plays an important role in hematopoietic progenitor cell growth and differentiation. It is also expressed by mast cells, macrophages and a CD5+ B cell subset. This antibody has been reported to block the binding of 125I-IL-3 to high and low affinity IL-3 receptors. In functional experiments, this antibody was found to inhibit acute myeloid leukemia cell proliferation, basophil histamine release, endothelial cell-mediated IL-8 secretion, and neutrophil transmigration. This antibody has been reported to be useful for immunoprecipitation, Western blot and immunofluorescent staining for flow cytometry. At the Fifth HLDA Workshop, the human IL-3 receptor was designated CD123.
The antibody was conjugated to BD Horizon™ BV650 which is part of the BD Horizon™ Brilliant Violet™ family of dyes. This dye is a tandem fluorochrome of BD Horizon™ BV421 with an Ex Max of 405-nm and an acceptor dye with an Em Max at 650-nm. BD Horizon™ BV650 can be excited by the violet laser and detected in a filter used to detect APC-like dyes (eg, 660/20-nm filter). Due to the excitation and emission characteristics of the acceptor dye, there will be spillover into the APC and Alexa Fluor® 700 detectors. However, the spillover can be corrected through compensation as with any other dye combination.
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