BUV496 Mouse Anti-Human CD16

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BD Horizon™ BUV496 Mouse Anti-Human CD16

Name: BUV496 Mouse Anti-Human CD16
Brand: BD Horizon™
SKU: 564653

Clone 3G8

(RUO)

Two-color flow cytometric analysis of human CD16 expression on human peripheral blood cells. Human peripheral blood cells were stained with APC Mouse Anti-Human CD56 antibody (Cat. No. 555518) and either BD Horizon™ BUV496 Mouse IgG1, κ Isotype Control (Cat. No. 564650; Left Panel) or BD Horizon BUV496 Mouse Anti-Human CD16 antibody (Cat. No. 564653/564654; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-color flow cytometric dot plots show the correlated expression patterns of CD16 versus CD56 (or Ig Isotype control staining) for gated events with the forward and side light-scatter characteristics of intact peripheral blood lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

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Product Details

Brand: BD Horizon™

Alternative Name: CD16;CD16A;FCGR3A;FcγRIIIA;FcRIIIa;CD16B;FCGR3B;FcγRIIIB;FcRIIIb

Reactivity: Human (QC Testing), Rhesus,Cynomolgus,Baboon (Tested in Development)

Isotype: Mouse BALB/c x DBA/2, also known as CD2F1 or CDF1 IgG1, κ

Immunogen: Human polymorphonuclear leukocytes

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Workshop Number: IV N409; V MR5, NK80

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

Regulatory Status: 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 BUV496 under optimum conditions, and unconjugated antibody and free BD Horizon BUV496 were removed.

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 of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).

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.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
BD Horizon Brilliant Ultraviolet 496 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; and 8,354,239.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
BD Horizon Brilliant Ultraviolet 496 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
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.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

Lysing Buffer RUO

Size 100 mL Cat No. 555899

APC Mouse Anti-Human CD56 (NCAM-1) RUO

Size 100 Tests Cat No. 555518

View Details

564653 Rev. 4

Antibody Details

3G8

The 3G8 monoclonal antibody specifically recognizes CD16a and CD16b, low affinity receptors for the Fc region of IgG. CD16a is ~50-65 kDa type I transmembrane glycoprotein that is encoded by FCGR3A (Fc fragment of IgG receptor IIIa) which belongs to the immunoglobulin superfamily. CD16a is also known as Fc-gamma RIII-alpha (Fc-gamma RIIIa or FcγRIIIA) or FcRIIIa and is expressed on natural killer cells, activated monocytes, macrophages, γδ T cells, immature thymocytes, and mast cells. CD16a binds immune-complexed or aggregated IgG and associates with CD247/TCRζ in NK cells and FcεRIγ chains in phagocytes and mast cells to transduce intracellular signals. CD16a functions in antibody-dependent cellular cytotoxicity (ADCC) and other antibody-dependent responses including phagocytosis, cytokine production or mediator release. CD16b is a ~48 kDa glycophosyl-phosphatidylinositol (GPI)-linked form that is encoded by FCGR3B (Fc fragment of IgG receptor IIIb). CD16b is also known as Fc-gamma RIII-beta (Fc-gamma RIIIb or FcγRIIIB) or FcRIIIb and is expressed on neutrophils and activated eosinophils. The extracellular region of CD16b is highly homologous to CD16a. CD16b also serves as a receptor for the Fc region of IgG and can bind immune-complexed or aggregated IgG and may be involved in neutrophil adhesion.

The 3G8 antibody also crossreacts with a subset of peripheral blood lymphocytes and monocytes, but not granulocytes, of baboon, rhesus, and cynomolgus monkeys. Multicolor analysis reveals that the distribution on lymphocytes is similar to that found in human studies with the majority of CD16-positive lymphocytes being both CD3 and CD20 negative.

This clone also cross-reacts with a subset of peripheral blood lymphocytes and monocytes, but not granulocytes, of baboon and both rhesus and cynomologus macaque monkeys. Multi-color analysis reveals that the distribution on lymphocytes is similar to that found in human studies with the majority of CD16-positive lymphocytes being both CD3 and CD20 negative.

The antibody was conjugated to BD Horizon BUV496 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 496-nm. BD Horizon BUV496 can be excited by the ultraviolet laser (355 nm) and detected with a 515/30 nm filter with a 450LP. Due to the excitation of the acceptor dye by other laser lines, there may be significant spillover into the channel detecting BD Horizon V500 or BV510 (e.g., 525/40-nm filter). However, the spillover can be corrected through compensation as with any other dye combination.

564653 Rev. 4

Format Details

BUV496

The BD Horizon Brilliant™ Ultraviolet 496 (BUV496) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 496-nm. BUV496, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 500-nm (e.g., 515/30-nm bandpass filter). The acceptor dye can be excited by the Violet (405-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BUV496

Excitation Source: Ultraviolet 355 nm

Excitation Max: 350 nm

Emission Max: 496 nm

564653 Rev.4

Citations & References

Development References (9)

Fleit HB, Wright SD, Durie CJ, Valinsky JE, Unkeless JC. Ontogeny of Fc receptors and complement receptor (CR3) during human myeloid differentiation. J Clin Invest. 1984; 73(2):516-525. (Clone-specific: Flow cytometry, Fluorescence microscopy, Immunofluorescence, Immunoprecipitation, Radioimmunoassay). View Reference
Fleit HB, Wright SD, Unkeless JC. Human neutrophil Fc gamma receptor distribution and structure. Proc Natl Acad Sci U S A. 1982; 79(10):3275-3279. (Immunogen: Blocking, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
Perussia B, Trinchieri G, Jackson A, et al. The Fc receptor for IgG on human natural killer cells: phenotypic, functional, and comparative studies with monoclonal antibodies. J Immunol. 1984; 133(1):180-189. (Clone-specific: Flow cytometry, Functional assay, Inhibition). View Reference
Schmidt RE. Non-lineage/natural killer section report: new and previously defined clusters. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:517-542.
Stroncek DF, Skubitz KM, Plachta LB, et al. Alloimmune neonatal neutropenia due to an antibody to the neutrophil Fc-gamma receptor III with maternal deficiency of CD16 antigen. Blood. 1991; 77(7):1572-1580. (Clone-specific: Immunofluorescence, Immunoprecipitation). View Reference
Vossebeld PJ, Homburg CH, Roos D, Verhoeven AJ. The anti-Fc gamma RIII mAb 3G8 induces neutrophil activation via a cooperative actin of Fc gamma RIIIb and Fc gamma RIIa. Int J Biochem Cell Biol. 1997; 29(3):465-473. (Clone-specific: Activation, Functional assay). View Reference
Wirthmueller U, Kurosaki T, Murakami MS, Ravetch JV. Signal transduction by Fc gamma RIII (CD16) is mediated through the gamma chain. J Exp Med. 1992; 175(5):1381-1390. (Clone-specific: Activation, Functional assay). View Reference
Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.

View All (9)

564653 Rev. 4

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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.