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Multiparameter flow cytometric analysis of CD24 expression on human peripheral blood leucocyte populations. Human whole blood was stained with FITC Mouse Anti-Human CD19 (Cat. No. 555412/560994) and either BD Horizon™ BB700 Mouse IgG2a, κ Isotype Control (Cat. No. 566419; Left Plot) or BD Horizon BB700 Mouse Anti-Human CD24 antibody (Cat. No. 566524/566525; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-parameter flow cytometric contour plots showing the correlated expression of CD24 (or Ig Isotype control staining) versus CD19 signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Horizon™ BB700 Mouse Anti-Human CD24
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
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 for the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.
For optimal results, it is recommended to perform two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.
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.
- 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.
- The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. Diagnostic uses require a separate license.
- 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.
- BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The ML5 monoclonal antibody specifically binds to CD24 which is also known as CD24A, signal transducer CD24, small cell lung carcinoma cluster 4 antigen, or BA-1. CD24 is a 35-70 kDa glycophosphatidylinositol (GPI)-linked glycoprotein whose glycosylation pattern is highly variable and cell-type dependent. CD24 is expressed on neutrophils, eosinophils, dendritic cells, neural cells, epithelial cells, muscle cells, and many types of cancer cells. CD24 functions as an adhesion receptor. Several different ligands have been identified for CD24 including CD62P (P-selectin) which is expressed on activated platelets and activated endothelium. CD24 is variably expressed on all B lineage cells, except plasma cells, and can play a role in regulating the activation, proliferation or differentiation of these cells.
The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.
Development References (8)
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Chtanova T, Tangye SG, Newton R, et al. T follicular helper cells express a distinctive transcriptional profile, reflecting their role as non-Th1/Th2 effector cells that provide help for B cells. J Immunol. 2004; 173(1):68-78. (Clone-specific: Flow cytometry). View Reference
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Fang X, Zheng P, Tang J, Liu Y. CD24: from A to Z.. Cell Mol Immunol. 2010; 7(2):100-3. (Biology). View Reference
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Le Bien TW. CD24 Workshop Panel 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; 1995:539-541.
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McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
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Pirruccello SJ, Bicak MS. Biochemical characterization of the CD24 antibody panel and cell-cycle studies of CD24 antigen expression on pre-B-ALL cell lines. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:83-84.
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Salamone MC, Rosselot C, Salamone GV, Barboza M, Kado M, Fainboim L. Antibodies recognizing CD24 LAP epitope on human T cells enhance CD28 and IL-2 T cell proliferation . J Leukoc Biol. 2001; 69(2):215-223. (Clone-specific: Flow cytometry). View Reference
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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.
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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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.