Flow cytometric analysis of CD15 expression on human peripheral blood granulocytes. Whole blood was stained with PE-Cy™7 Mouse Anti-Human CD15 (Cat. No. 560827; solid line histogram) or with a PE-Cy™7 Mouse IgM, κ Isotype Control (Cat. No. 560856; dashed line histogram). The erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable granulocytes. Flow cytometry was performed on a BD™ LSR II.
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Flow cytometric analysis of CD15 expression on human peripheral blood granulocytes. Whole blood was stained with PE-Cy™7 Mouse Anti-Human CD15 (Cat. No. 560827; solid line histogram) or with a PE-Cy™7 Mouse IgM, κ Isotype Control (Cat. No. 560856; dashed line histogram). The erythrocytes were lysed with Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from gated events with the forward and side light-scattering characteristics of viable granulocytes. Flow cytometry was performed on a BD™ LSR II.
Product Details
BD Pharmingen™
Lewis X; Le-X; X-Hapten; SSEA-1; 3-FAL
Human (QC Testing)
Mouse IgM, κ
Human Mononuclear Cells
Flow cytometry (Routinely Tested), Immunohistochemistry-paraffin (Tested During Development)
5 µl
IV M141
AB_10563901
Aqueous buffered solution containing BSA, protein stabilizer, 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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.
Product Notices
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Cy is a trademark of GE Healthcare.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
- Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE-Cy™7 Mouse IgM, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560856
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560827 Rev. 3
Antibody Details
HI98
The HI98 monoclonal antibody specifically reacts with 3-fucosyl-N-acetyllactosamine (3-FAL), a 220 kDa carbohydrate structure, also called X-hapten, SSEA-1, CD15 or Lewis X. This structure is found on a variety of cell surface glycolipids and glycoproteins. 3-FAL is expressed on >95% of granulocytes, including neutrophils and eosinophils, and to a varying degree on monocytes, but not on lymphocytes or basophils. CD15 plays a role in mediating phagocytosis, bactericidal activity and chemotaxis. This antibody is also suitable for staining formalin-fixed, paraffin-embedded tissue sections without pretreatment. Since the Abs are recognizing a carbohydrate epitope (3-fucosyl-N-acetyllactosamine) they also should work across species and not only for human.
560827 Rev. 3
Format Details
PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-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.
PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560827 Rev.3
Citations & References
View product citations for antibody "560827" on CiteAb
Development References (3)
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Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
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Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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Lund-Johansen F, Olweus J, Horejsi V, et al. Activation of human phagocytes through carbohydrate antigens (CD15, sialyl-CD15, CDw17, and CDw65).. J Immunol. 1992; 148(10):3221-9. (Biology). View Reference
560827 Rev. 3
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.