Multiparameter flow cytometric analysis of CD66b expression on human peripheral blood leucocyte populations. Whole blood was stained with either Biotin Mouse IgM, κ Isotype Control (Left Plot; Cat. No. 553473) or Biotin Mouse Anti-Human CD66b antibody (Right Plot; Cat. No. 567941) at 0.06 µg/test, followed by PE Streptavidin (Cat. No. 554061). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing CD66b expression (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) were derived from gated events with the side and forward light-scattering characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of CD66b expression on human peripheral blood leucocyte populations. Whole blood was stained with either Biotin Mouse IgM, κ Isotype Control (Left Plot; Cat. No. 553473) or Biotin Mouse Anti-Human CD66b antibody (Right Plot; Cat. No. 567941) at 0.06 µg/test, followed by PE Streptavidin (Cat. No. 554061). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing CD66b expression (or Ig Isotype control staining) versus side light-scatter signals (SSC-A) were derived from gated events with the side and forward light-scattering characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CEACAM8; CGM6; NCA-95
Human (QC Testing)
Mouse BALB/c IgM, κ
Human Granulocytes
Flow cytometry (Routinely Tested)
0.5 mg/ml
V 5T-127, MA020
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 with biotin under optimum conditions, and unreacted biotin was removed. Store undiluted at 4°C.
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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567941 Rev. 1
Antibody Details
G10F5
The G10F5 monoclonal antibody specifically binds to CD66b, also known as Carcinoembryonic antigen-related cell adhesion molecule 8 (CEACAM8). CD66b is a glycosylphosphatidylinositol (GPI) linked protein with a molecular weight of 100 kDa expressed on granulocytes. This molecule was previously clustered as CD67 in the Fourth Human Leucocyte Differentiation Antigen (HLDA) Workshop and renamed CD66b in the Fifth HLDA Workshop. CD66b is a member of the carcinoembryonic antigen (CEA)-like glycoprotein family present on granulocytes and referred to as non-specific crossreacting antigens (NCA). Granulocyte activation induced with soluble stimulators (calcium ionophore, phorbol myristate acetate, N-formylmethionyl- leucyl-phenylalanine) results in release and increased expression of NCA. Findings suggest that these molecules may play a role in phagocytosis, chemotaxis and adherence.
567941 Rev. 1
Format Details
Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
567941 Rev.1
Citations & References
Development References (7)
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Hemler ME, Kassner P, Bodorova J. CD66 and CD67 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; 1995:889-899.
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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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Kuijpers TW, van der Schoot CE, Hoogerwerf M, Roos D. Cross-linking of the carcinoembryonic antigen-like glycoproteins CD66 and CD67 induces neutrophil aggregation. J Immunol. 1993; 151(9):4934-4940. (Biology). View Reference
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Kuroki M, Matsuo Y, Kinugasa T, Matsuoka Y. Augmented expression and release of nonspecific cross-reacting antigens (NCAs), members of the CEA family, by human neutrophils during cell activation. J Leukoc Biol. 1992; 52(5):551-557. (Biology). View Reference
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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. (Clone-specific: Blocking, 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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Thompson JS, Brown SA, Rhoades JL, Burch J, Oberle EM. G10F5 (Workshop no. 310) reacts with a Pronase resistant epitope whose tissue distribution differs from CD15 monoclonal antibodies. In: McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:713-714.
567941 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.
