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BV711 Mouse Anti-Human IL-3Rα (CD123)
Product Details
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BD OptiBuild™
IL3RA; IL3R; IL-3R-alpha; IL-3RA
Human (Tested in Development)
Mouse BALB/c IgG1, κ
Human IL3RA Transfected Cell Line
Flow cytometry (Qualified)
0.2 mg/ml
AB_2875804
Aqueous buffered solution containing ≤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 the dye under optimum conditions that minimize unconjugated dye and antibody.

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. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
  10. BD Horizon Brilliant Violet 711 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
  11. Cy is a trademark of GE Healthcare.
751833 Rev. 1
Antibody Details
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6H6

The 6H6 monoclonal antibody specifically recognizes the Interleukin-3 receptor alpha chain (IL-3Ra) which is also known as CD123. IL-3Ra (CD123) is a ~70 kDa type I transmembrane glycoprotein that is encoded by IL3RA (interleukin 3 receptor subunit alpha) which belongs to the type I cytokine receptor family within the immunoglobulin gene superfamily. This receptor chain consists of an extracellular region that contains an immunoglobulin-like N-terminal domain (NTD) with a fibronectin type III (FnIII) fold followed by two more FnIII domains that form the cytokine receptor module (CRM), a transmembrane region, and an intracellular tail. IL-3Ra (CD123) binds IL-3 specifically and with low affinity. IL-3Ra (CD123) forms a high-affinity signaling receptor for IL-3 (IL-3R) with the ß common chain (ßc; also known as, CD131) that is shared with the heterodimeric IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) receptors. IL-3Ra (CD123) is variably expressed on certain hematopoietic progenitor cells, basophils, eosinophils, mast cells, monocytes, macrophages, dendritic cells, megakaryocytes, and on some B cells, endothelial cells, and leukemia cells. Bound IL-3 can signal through IL-3R to promote the activation, proliferation, differentiation, and viability of these cells. Amongst monoclonal antibodies specific for human IL-3Ra (CD123), the 6H6 and 9F5 antibodies do not block IL-3 binding to the IL-3R whereas the 7G3 antibody does block IL-3 binding to its receptor in a dose-dependent manner.

The antibody was conjugated to BD Horizon™ BV711 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 711-nm.  BD Horizon BV711 can be excited by the violet laser and detected in a filter used to detect Cy™5.5 / Alexa Fluor® 700-like dyes (eg, 712/20-nm filter).  Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy5.5 detectors.  However, the spillover can be corrected through compensation as with any other dye combination.

751833 Rev. 1
Format Details
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BV711
The BD Horizon Brilliant Violet™ 711 (BV711) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This tandem fluorochrome is comprised of a BV421 donor with an excitation maximum (Ex Max) of 407-nm and an acceptor dye with an emission maximum (Em Max) at 713-nm. BV711, driven by BD innovation, is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 710-nm (e.g., a 712/20-nm bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV711
Violet 405 nm
407 nm
713 nm
751833 Rev.1
Citations & References
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Development References (6)

  1. Broughton SE, Hercus TR, Hardy MP, et al. Dual mechanism of interleukin-3 receptor blockade by an anti-cancer antibody.. Cell Rep. 2014; 8(2):410-9. (Biology). View Reference
  2. 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
  3. Miyajima A. CDw123 (Interleukin 3 receptor α chain) Workshop Panel report. In: 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.; 1997:854-855.
  4. 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. (Immunogen: Flow cytometry, Immunoprecipitation, Western blot). View Reference
  5. Yamada T, Sun Q, Zeibecoglou K, et al. IL-3, IL-5, granulocyte-macrophage colony-stimulating factor receptor alpha-subunit, and common beta-subunit expression by peripheral leukocytes and blood dendritic cells.. J Allergy Clin Immunol. 1998; 101(5):677-82. (Clone-specific: Flow cytometry). View Reference
  6. Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
View All (6) View Less
751833 Rev. 1

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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.