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      RB705 Mouse Anti-Human CD33
      RB705 Mouse Anti-Human CD33

      Multiparameter flow cytometric analysis of CD33 expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leukocytes were then stained with either BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat. No. 570261; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD33 antibody (Cat. No. 570641/570727; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD33 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

      RB705 Mouse Anti-Human CD33

      Multiparameter flow cytometric analysis of CD33 expression on Human peripheral blood leukocyte populations.  Human whole blood was first treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). The leukocytes were then stained with either BD Horizon™ RB705 Mouse IgG1, κ Isotype Control (Cat. No. 570261; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD33 antibody (Cat. No. 570641/570727; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD33 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

      Product Details
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      BD Horizon™
      Siglec-3; SIGLEC3; Sialic acid-binding Ig-like lectin 3; p67; gp67; My9
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Acute Myeloid Leukemia Blasts
      Flow cytometry (Routinely Tested)
      5 µl/test
      IV M505
      945
      AB_3685919
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

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

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      Product Notices

      1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. 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).
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
      6. 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.
      7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
      9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      11. For U.S. patents that may apply, see bd.com/patents.
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      570641 Rev. 1
      Antibody Details
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      WM53

      The WM53 monoclonal antibody specifically recognizes CD33 which is also known as Sialic acid-binding Ig-like lectin 3 (Siglec-3) or gp67. CD33 is a 67 kDa type I transmembrane glycoprotein that is variably expressed on myeloid progenitors, monocytes, macrophages, dendritic cells, neutrophils, basophils, mast cells, and on some activated T cells and NK cells. Normal lymphocytes, platelets, erythrocytes and pluripotent hematopoietic stem cells do not express the CD33 antigen. This glycoprotein reportedly functions as a sialic acid-dependent cell adhesion molecule and this function can be modulated by endogenous sialoglycoconjugates when CD33 is expressed on the membrane.

      570641 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      570641 Rev.1
      Citations & References
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      View product citations for antibody "570641" on CiteAb

      Development References (7)

      1. Favaloro EJ, Bradstock KF, Kabral A, Grimsley P, Berndt MC. Characterization of monoclonal antibodies to the human myeloid-differentiation antigen, 'gp67' (CD-33). Dis Markers. 1987; 5(4):215-225. (Immunogen: Blocking, Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunoprecipitation, Radioimmunoassay). View Reference
      2. Favaloro EJ, Bradstock KF, Kabral A, Grimsley P, Zowtyj H, Zola H. Further characterization of human myeloid antigens (gp160,95; gp150; gp67): investigation of epitopic heterogeneity and non-haemopoietic distribution using panels of monoclonal antibodies belonging to CD-11b, CD-13 and CD-33. Br J Haematol. 1988; 69(2):163-171. (Clone-specific: Blocking, Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunohistochemistry, Radioimmunoassay). View Reference
      3. Favaloro EJ, Moraitis N, Koutts J, Exner T, Bradstock KF. Endothelial cells and normal circulating haemopoietic cells share a number of surface antigens. Thromb Haemost. 1989; 61(2):217-224. (Clone-specific: ELISA). View Reference
      4. Freeman SD, Kelm S, Barber EK, Crocker PR. Characterization of CD33 as a new member of the sialoadhesin family of cellular interaction molecules. Blood. 1995; 85(8):2005-2012. (Biology). View Reference
      5. Hernández-Caselles T, Martínez-Esparza M, Pérez-Oliva AB, et al. A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing.. J Leukoc Biol. 2006; 79(1):46-58. (Clone-specific: Flow cytometry). View Reference
      6. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
      7. Nakamura Y, Noma M, Kidokoro M, et al. Expression of CD33 antigen on normal human activated T lymphocytes.. Blood. 1994; 83(5):1442-3. (Biology). View Reference
      View All (7) View Less
      570641 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.