RB705 Mouse Anti-Human CD66b

BD Horizon™ RB705 Mouse Anti-Human CD66b

Name: RB705 Mouse Anti-Human CD66b
Brand: RB705 Mouse Anti-Human CD66b
SKU: 570599

Clone G10F5 (RUO)

Multiparameter flow cytometric analysis of CD66b expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either no BD Horizon™ RB705 conjugated antibody (Autofluorescence control; Left Plot) or BD Horizon™ RB705 Mouse Anti-Human CD66b antibody (Cat. No. 570599/570687; Right Plot). The bivariate pseudocolor density plot showing CD66b expression (or cellular Autofluorescence) versus side light-scatter signals (SSC-A) were derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software.

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Product Details

Brand: BD Horizon™

Alternative Name: CEACAM8; CGM6; NCA-95

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgM, κ

Immunogen: Human Granulocytes

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Workshop Number: V 5T-127, MA020

Entrez Gene ID: 1088

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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

When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
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).
Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
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.
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Human BD Fc Block™ RUO

Size 50 µg Cat No. 564219

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

570599 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.

570599 Rev. 1

Format Details

RB705

The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.

Format: RB705

Excitation Source: Blue 488 nm

Excitation Max: 498 nm

Emission Max: 707 nm

570599 Rev.1

Citations & References

View product citations for antibody "570599" on CiteAb

Development References (7)

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.
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
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
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
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
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.
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)

570599 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.

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.