Skip to main content Skip to navigation
BB515 Mouse Anti-Human CD66b
BB515 Mouse Anti-Human CD66b

Flow cytometric analysis of CD66b expression on human peripheral blood granulocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with either BD Horizon BB515 Mouse IgM, κ Isotype Control (Cat. No. 564680; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD66b antibody (Cat. No. 564679; bold solid line histogram). Alternatively, the cells were stained with FITC Mouse Anti-Human CD66b antibody (Cat. No. 555724/561927; thin solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202).

       Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD66b antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD66b antibody versus FITC Anti-CD66b antibody (Right Panel). The fluorescence histograms showing CD66b expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact granulocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD66b expression on human peripheral blood granulocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with either BD Horizon BB515 Mouse IgM, κ Isotype Control (Cat. No. 564680; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human CD66b antibody (Cat. No. 564679; bold solid line histogram). Alternatively, the cells were stained with FITC Mouse Anti-Human CD66b antibody (Cat. No. 555724/561927; thin solid line histogram). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202).

       Overlaid histograms are shown to facilitate staining comparisons between: BB515 Anti-CD66b antibody versus its Ig Isotype Control (Left Panel), and BB515 Anti-CD66b antibody versus FITC Anti-CD66b antibody (Right Panel). The fluorescence histograms showing CD66b expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of intact granulocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Product Details
Down Arrow Up Arrow


BD Horizon™
CEACAM8; CGM6; NCA-95
Human (QC Testing)
Mouse BALB/c IgM, κ
Human Granulocytes
Flow cytometry (Routinely Tested)
5 µl
V 5T-127, MA020
AB_2738889
Aqueous buffered solution containing ≤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 BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

For optimal results, it is recommended to perform 2 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

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564679 Rev. 1
Antibody Details
Down Arrow Up Arrow
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.

The antibody was conjugated to BD Horizon BB515 which was developed exclusively by BD Biosciences. With an excitation max of 490 nm and an emission max of 515 nm, BD Horizon BB515 can be excited by the 488 nm laser and detected in a standard FITC set (eg, 530/30-nm filter). This dye provides a much brighter alternative to FITC with less spillover into the PE detector.

564679 Rev. 1
Format Details
Down Arrow Up Arrow
BB515
The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
BB515
Blue 488 nm
490 nm
515 nm
564679 Rev.1
Citations & References
Down Arrow Up Arrow

Development References (7)

  1. 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.
  2. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  3. 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
  4. 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
  5. 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
  6. 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.
  7. 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.
View All (7) View Less
564679 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.