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BD Horizon™ BUV395 Mouse Anti-Human CD16
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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563784/563785; 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.
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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563784/563785; 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.
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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563784/563785; 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.
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™ BUV395 Mouse IgG1, κ Isotype Control (Cat. No. 563547; Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human CD16 antibody (Cat. No. 563784/563785; 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.
Product Details
BD Horizon™
CD16;CD16A;FCGR3A;FcγRIIIA;FcRIIIa;CD16B;FCGR3B;FcγRIIIB;FcRIIIb
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c x DBA/2, also known as CD2F1 or CDF1 IgG1, κ
Human polymorphonuclear leukocytes
Flow cytometry (Routinely Tested)
5 µl
IV N409; V MR5, NK80
AB_2744293
Aqueous buffered solution containing BSA and ≤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 BUV395 under optimum conditions, and unconjugated antibody and free BD Horizon BUV395 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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.
- BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 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
BUV395 Mouse Anti-Human CD16 RUO
Size
25 Tests
Cat No.
563784
563785 Rev. 3
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™ BUV395 which has been exclusively developed by BD Biosciences as an optimal dye for use on a 355 nm laser equipped instrument. With an Ex Max at 348 nm and an Em Max at 395 nm, this dye has virtually no spillover into any other detector. BD Horizon™ BUV395 can be excited with a 355 nm laser and detected with a 379/28 filter.
563785 Rev. 3
Format Details
BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
BUV395
348 nm
395 nm
563785 Rev.3
Citations & References
View product citations for antibody "563785" on CiteAb
Development References (8)
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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
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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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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
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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.
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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
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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
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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
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
563785 Rev. 3
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.