Multiparameter flow cytometric analysis of CD15 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). 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 CD15 antibody (Cat. No. 570583/570671; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD15 (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.
Multiparameter flow cytometric analysis of CD15 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). 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 CD15 antibody (Cat. No. 570583/570671; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD15 (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
BD Horizon™
LeX; Lewis X; SSEA-1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Polymorphonuclear Leucocytes
Flow cytometry (Routinely Tested)
5 µl/test
V MA088
2526
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.
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.
Product Notices
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
RB705 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570261
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Human BD Fc Block™ RUO
Size
50 µg
Cat No.
564219
570583 Rev. 1
Antibody Details
7C3.rMAb
The 7C3.rMAb is a recombinant monoclonal antibody that specifically recognizes human CD15 which is a terminal carbohydrate epitope, 3-fucosyl-N-acetyllactosamine (3-FAL), on cell surface glycoproteins or glycolipids. CD15 is also known as Lewis X (LeX), X-Hapten, or Stage-Specific Embryonic Antigen-1 (SSEA-1). The 7C3.rMAb was derived from the 7C3 (also known as, PMN7C3) hybridoma. The 7C3 antibody was validated at HLDA V (Workshop Number MA88). The 7C3.rMAb has the same IgV-region heavy chain domain and Ig, κ light chain as the original mouse 7C3 IgG3, κ antibody. The remaining Ig constant heavy chain of 7C3.rMAb is derived from the mouse IgG1 heavy chain. CD15 is expressed on a variety of cell types including neutrophils, eosinophils, monocytes, macrophages, mast cells, and Langerhans cells. CD15 is also expressed on some epithelial cells, activated lymphocytes as well as tumor cells. CD15 is not expressed by platelets or erythrocytes. CD15 plays various roles in mediating cellular adhesion, activation, migration, and phagocytosis.
570583 Rev. 1
Format Details
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.
RB705
Blue 488 nm
498 nm
707 nm
570583 Rev.1
Citations & References
View product citations for antibody "570583" on CiteAb
Development References (8)
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Ball ED, Persichetti J, Roscoe R, Nimgaonkar M, Winkelstein A. Expression of CD15 on CD34+ cells of the bone marrow, peripheral blood, and umbilical cord blood. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:795-798.
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Ball ED. CD15 cluster workshop report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:790-794.
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Larsen GR, Sako D, Ahern TJ, et al. P-selectin and E-selectin. Distinct but overlapping leukocyte ligand specificities.. J Biol Chem. 1992; 267(16):11104-10. (Clone-specific: Cell separation). View Reference
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Masat T, Feliu E, Villamor N, et al. Immunophenotypic and ultrastructural study in peripheral blood neutrophil granulocytes following bone marrow transplantation.. Br J Haematol. 1997; 98(2):299-307. (Clone-specific: Flow cytometry). View Reference
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Nauseef WM, Root RK, Newman SL, Malech HL. Inhibition of zymosan activation of human neutrophil oxidative metabolism by a mouse monoclonal antibody.. Blood. 1983; 62(3):635-44. (Immunogen: Functional assay, Inhibition). View Reference
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Phillips ML, Schwartz BR, Etzioni A, et al. Neutrophil adhesion in leukocyte adhesion deficiency syndrome type 2.. J Clin Invest. 1995; 96(6):2898-906. (Clone-specific: Flow cytometry). View Reference
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Sedlak J, Chorvath B, Hunakova L, Bizik J, Karpatova M, Duraj J. CD15 (Lewisx) antigen: expression on non-hemopoietic cell lines and modulation on retinois-acid-induced HL-60 cells (analysedwith CD15 Panel mAb). In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:798-800.
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de Almeida LY, Pereira-Martins DA, Weinhäuser I, et al. The Combination of Gefitinib With ATRA and ATO Induces Myeloid Differentiation in Acute Promyelocytic Leukemia Resistant Cells.. Front Oncol. 2021; 11:686445. (Clone-specific: Flow cytometry). View Reference
570583 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.