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Profile of peripheral blood granulocytes analyzed by flow cytometry
Multivariate flow cytometric analysis of CD66c expression on human peripheral blood leucocyte populations. Human whole blood was stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 555749; Left Plot) or PE Mouse Anti-Human CD66c antibody (Cat. No. 551478; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing CD66c expression (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals were derived from events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
BD Pharmingen™ PE Mouse Anti-Human CD66c
BD Pharmingen™ PE Mouse Anti-Human CD66c
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
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
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The B6.2 monoclonal antibody reacts with a glycosylphosphatidylinositol-anchored glycoprotein present on granulocytes. Antibody B6.2 was studied as recognizing CD66c in the VI Human Leukocyte Differentiation Antigen workshop. CD66 antigens also known as the carcinoembryonic antigen (CEA) family of molecules, are closely related to the immunoglobulin super family of glycoproteins. Studies on CD66 molecules suggest a potential adhesion function in vivo. These molecules exhibit both homophilic and heterophilic adhesion. CEA family members may be involved in transmembrane signalling and activation of neutrophils. This clone has been found to be N-terminal domain reactive, reacted preferentially with the native protein and were conformationally dependent.
Development References (7)
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Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
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Kuroki M, Arakawa F, Matsuo Y, et al. Molecular cloning of nonspecific cross-reacting antigens in human granulocytes. J Biol Chem. 1991; 266(18):11810-11817. (Biology). 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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Skubitz KM, Campbell KD, Ahmed K, Skubitz AP. CD66 family members are associated with tyrosine kinase activity in human neutrophils. J Immunol. 1995; 155(11):5382-5390. (Biology). View Reference
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Szpak CA, Johnston WW, Lottich SC, Kufe D, Thor A, Schlom J. Patterns of reactivity of four novel monoclonal antibodies (B72.3, DF3, B1.1 and B6.2) with cells in human malignant and benign effusions. Acta Cytol. 1984; 28(4):356-367. (Biology). View Reference
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Thompson JA, Grunert F, Zimmermann W. Carcinoembryonic antigen gene family: molecular biology and clinical perspectives. J Clin Lab Anal. 1991; 5(5):344-366. (Biology). View Reference
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Watt SM, Teixeira AM, Zhou GQ, et al. Homophilic adhesion of human CEACAM1 involves N-terminal domain interactions: structural analysis of the binding site. Blood. 2001; 98(5):1469-1479. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.