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BD Horizon™ V500 Mouse Anti-Human CD15
Clone HI98 (also known as HIM1) (RUO)
Flow cytometric analysis of CD15 expression on human peripheral blood granulocytes. Whole blood was stained with BD Horizon™ V500 Mouse Anti-Human CD15 antibody (Cat. No. 561585; solid line histogram) or with a BD Horizon™ V500 Mouse IgM, κ Isotype Control (Cat. No. 561574; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD15 expression on human peripheral blood granulocytes. Whole blood was stained with BD Horizon™ V500 Mouse Anti-Human CD15 antibody (Cat. No. 561585; solid line histogram) or with a BD Horizon™ V500 Mouse IgM, κ Isotype Control (Cat. No. 561574; dashed line histogram). The erythrocytes were lysed with BD PharmLyse™ Lysing Buffer (Cat. No. 555899). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable granulocytes. Flow cytometry was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
Lewis X; Le-X; X-Hapten; SSEA-1; 3-FAL
Human (QC Testing)
Mouse IgM, κ
Human Mononuclear Cells
Flow cytometry (Routinely Tested)
5 µl
IV M141
AB_10896278
Aqueous buffered solution containing protein stabilizer, glycerol and ≤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 with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- 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.
- 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.
- BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
Companion Products
Stain Buffer (FBS) RUO
Size
500 ML
Cat No.
554656
Lysing Buffer RUO
Size
100 ML
Cat No.
555899
V500 Mouse IgM, κ Isotype Control RUO
Size
0.1 mg
Cat No.
561574
561585 Rev. 1
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.
The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is
available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.
When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
561585 Rev. 1
Format Details
V500
BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.When compensating dyes in this spectral range (such as BD Horizon V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls. Due to spectral differences between cells and beads in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different V500 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.
V500
415 nm
499 nm
561585 Rev.1
Citations & References
View product citations for antibody "561585" on CiteAb
Development References (3)
-
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.
-
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
561585 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.