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Flow cytometric analysis of FcγRIV (CD16-2) expression on mouse bone marrow and peripheral blood cells. BALB/c mouse bone marrow cells and peripheral blood were treated separately with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The cells were washed and then stained with either PE Hamster IgG2, κ Isotype Control (Cat. No. 550085) or PE Hamster Anti-Mouse FcγRIV antibody (Cat. No. 565615). In addition, the bone marrow cells were stained with Alexa Fluor® 647 Anti-CD11b antibody (Cat. No. 557686), whereas the peripheral blood cells were stained with Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669). Two-color flow cytometric contour plots showing the correlated expression of FcγRIV (CD16-2) [or Ig Isotype control staining] versus CD11b (Panel 1; Bone Marrow Cells) or CD45R/B220 (Panel 2; Peripheral Blood Leucocytes) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
BD Pharmingen™ PE Armenian Hamster Anti-Mouse FcγRIV (CD16-2)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
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.
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
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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.