Two-color flow cytometric analysis of FcγRIV (CD16-2) on mouse bone marrow cells. BALB/c mouse bone marrow cells were preincubated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse red blood cells. The cells were washed and then stained with either Purified Hamster IgG2, κ Isotype Control (Cat. No. 559277; Left Plot) or Purified Armenian Hamster Anti-Mouse FcγRIV (CD16-2) antibody (Cat. No. 567208; Right Plot) at 0.25 µg/test. The cells were washed and then stained with Alexa Fluor® 647 Rat Anti-CD11b antibody (Cat. No 557686) and PE Mouse Anti-Armenian and Syrian Hamster IgG Cocktail (Cat. No. 554056). The two-color contour plot showing the correlated expression of FcγRIV (CD16-2) [or Ig Isotype Control staining] versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of FcγRIV (CD16-2) on mouse bone marrow cells. BALB/c mouse bone marrow cells were preincubated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse red blood cells. The cells were washed and then stained with either Purified Hamster IgG2, κ Isotype Control (Cat. No. 559277; Left Plot) or Purified Armenian Hamster Anti-Mouse FcγRIV (CD16-2) antibody (Cat. No. 567208; Right Plot) at 0.25 µg/test. The cells were washed and then stained with Alexa Fluor® 647 Rat Anti-CD11b antibody (Cat. No 557686) and PE Mouse Anti-Armenian and Syrian Hamster IgG Cocktail (Cat. No. 554056). The two-color contour plot showing the correlated expression of FcγRIV (CD16-2) [or Ig Isotype Control staining] versus CD11b was derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
CD16-2; Fcgr4; FcgRIV; Fc gammaRIV; FcgammaRIV; Fcgr3a; Fcrl3
Mouse (QC Testing)
Armenian Hamster IgG
Mouse FcγRIV Recombinant Protein
Flow cytometry (Routinely Tested)
0.5 mg/ml
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. Store undiluted at 4°C.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
Companion Products
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Purified Hamster IgG2, κ Isotype Control RUO
Size
0.5 mg
Cat No.
559277
Alexa Fluor® 647 Rat Anti-CD11b RUO
Size
0.1 mg
Cat No.
557686
PE Mouse Anti-Armenian and Syrian Hamster IgG Cocktail RUO
Size
0.2 mg
Cat No.
554056
567208 Rev. 2
Antibody Details
9E9
The 9E9 monoclonal antibody specifically binds to Fc receptor, IgG, low affinity IV (FcγRIV), which is also known as CD16-2. FcγRIV is encoded by Fcgr4 and belongs to the family of receptors for the Fc region of IgG (FcγRs), which also includes FcγRI (CD64), FcγRII (CD32), and FcγRIII (CD16) within the Ig superfamily. FcγRIV is expressed on monocytes, macrophages, dendritic cells, and neutrophils. FcγRIV, which serves as a ligand-biding subunit, requires the common Fcγ chain for expression and signaling. This complex serves as a cell activating receptor when bound by IgG2a or IgG2b. FcγRIV plays various roles in inflammation including neutrophil trafficking and mast cell degranulation. FcγRIV can also function as a low-affinity IgE receptor and promote IgE-induced inflammation. The 9E9 antibody can reportedly inhibit cellular FcγRIV (CD16-2) function and block non-antigen-specific binding of immune complexes to improve the quality of immunological assays.
567208 Rev. 2
Format Details
Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
567208 Rev.2
Citations & References
View product citations for antibody "567208" on CiteAb
Development References (5)
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Bruhns P. Properties of mouse and human IgG receptors and their contribution to disease model. Blood. 2012; 119(24):5640-5649. (Biology). View Reference
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Mancardi DA, Iannascoli B, Hoos S, England P, Daëron M, Bruhns P. FcgammaRIV is a mouse IgE receptor that resembles macrophage FcepsilonRI in humans and promotes IgE-induced lung inflammation. J Clin Invest. 2008; 118(11):3738-3750. (Clone-specific: Blocking, Flow cytometry, Functional assay). View Reference
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Nimmerjahn F, Lux A, Albert H, et al. FcγRIV deletion reveals its central role for IgG2a and IgG2b activity in vivo. Proc Natl Acad Sci U S A. 2010; 107(45):19396-19401. (Clone-specific: Blocking, Flow cytometry, Immunohistochemistry). View Reference
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Nimmerjahn F, Ravetch JV. FcγRs in health and disease. Curr Top Microbiol Immunol. 2011; 350:105-125. (Biology). View Reference
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Syed SN, Konrad S, Wiege K, et al. Both FcgammaRIV and FcgammaRIII are essential receptors mediating type II and type III autoimmune responses via FcRgamma-LAT-dependent generation of C5a. Eur J Immunol. 2009; 39(12):3343-3356. (Clone-specific: Flow cytometry). View Reference
567208 Rev. 2
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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.