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BV421 Mouse Anti-Human CD123
BV421 Mouse Anti-Human CD123
Multicolor flow cytometric analysis of CD123 expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV421 Mouse Anti-Human CD123 antibody (Cat. No. 562517/565928) and APC Mouse Anti-Human CD45RO antibody (Cat. No. 560899/559865). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two color flow cytometric dot plot shows the correlated expression of CD45RO versus CD123 for gated events with the forward and side light-scatter characteristics of viable lymphocytes. Quadrant markers were placed based upon the corresponding fluorescent immunoglobulin isotype controls (data not shown). Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Multicolor flow cytometric analysis of CD123 expression on human peripheral blood lymphocytes. Human whole blood was stained with the BD Horizon™ BV421 Mouse Anti-Human CD123 antibody (Cat. No. 562517/565928) and APC Mouse Anti-Human CD45RO antibody (Cat. No. 560899/559865). The erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two color flow cytometric dot plot shows the correlated expression of CD45RO versus CD123 for gated events with the forward and side light-scatter characteristics of viable lymphocytes. Quadrant markers were placed based upon the corresponding fluorescent immunoglobulin isotype controls (data not shown). Flow cytometry was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
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BD Horizon™
IL3RA; IL-3RA; IL-3Rα; IL-3R-alpha; Interleukin-3 receptor subunit alpha
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human IL-3Rα Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
VI C-67
3563
AB_11153668
Aqueous buffered solution containing BSA 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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
565928 Rev. 1
Antibody Details
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9F5

The 9F5 monoclonal antibody specifically binds to CD123. CD123 is the 70 kDa IL-3 receptor α chain (IL-3Rα) that associates with the 120-140 kDa β subunit (CD131/Common β-chain/βc) to form the functional IL-3 receptor complex. The βc chain is also shared with distinct α chain subunits to form the functional heterodimeric receptors for interleukins IL-5 and GM-CSF. IL-3Rα is expressed on a subset of peripheral blood dendritic cells, myeloid precursors, basophils, mast cells, macrophages, and megakaryocytes. Reports indicate that IL-3Rα is also expressed on lymphocytes. The IL-3R plays an important role in hematopoietic progenitor cell growth and differentiation. This antibody does not block binding of IL-3 to the IL-3 receptor.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific BlueTM conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

565928 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
565928 Rev.1
Citations & References
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Development References (5)

  1. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  2. Korpelainen EI, Gamble JR, Smith WB, et al. The receptor for interleukin 3 is selectively induced in human endothelial cells by tumor necrosis factor alpha and potentiates interleukin 8 secretion and neutrophil transmigration.. Proc Natl Acad Sci USA. 1993; 90(23):11137-41. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  3. Macardle PJ, Chen Z, Shih CY, et al. Characterization of human leucocytes bearing the IL-3 receptor. Cell Immunol. 1996; 168(1):59-68. (Biology). View Reference
  4. Smith WB, Guida L, Sun Q, et al. Neutrophils activated by granulocyte-macrophage colony-stimulating factor express receptors for interleukin-3 which mediate class II expression. Blood. 1995; 86(10):3938-3944. (Clone-specific: Flow cytometry). View Reference
  5. Sun Q, Woodcock JM, Rapoport A, et al. Monoclonal antibody 7G3 recognizes the N-terminal domain of the human interleukin-3 (IL-3) receptor alpha-chain and functions as a specific IL-3 receptor antagonist.. Blood. 1996; 87(1):83-92. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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565928 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.