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Flow cytometric analysis of FcεRIα expression on MC/9 cells. Cells from the mouse MC/9 (Mast Cells, ATCC CRL-8306) cell line were stained with PE Hamster IgG1, κ Isotype Control (Cat. No. 553972; dashed line histogram) or PE Mouse Anti-FcεRIα antibody (Cat. No. 566994/566995; solid line histogram) at 0.25 μg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. The fluorescence histogram showing FcεRIα expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable (7-AAD-negative) cells. 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.
Multiparameter flow cytometric analysis of FcεRIα expression on mouse spleen cells. C57BL/6 mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Hamster Anti-Mouse CD3e (Cat. No. 553062/553061/561827), FITC Rat anti-Mouse CD45R/B220 (Cat. No. 553088/553087/561877), APC Rat Anti-Mouse CD49b (Cat. No. 560628) antibodies and either PE Hamster IgG1, κ Isotype Control (Cat. No. 553972; Left Plot) or PE Hamster Anti-Mouse FcεRIα antibody (Cat. No. 566994/566995; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Two-parameter pseudocolor density plots showing the correlated expression of FcεRIα (or Ig Isotype control staining) versus CD49b were derived from CD3-negative B220-negative gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Hamster Anti-Mouse FcεR1α
BD Pharmingen™ PE Hamster Anti-Mouse FcεR1α
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
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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Product Notices
- 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.
- 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.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The MAR-1 monoclonal antibody specifically recognizes Fc-epsilon RI-alpha (FceR1 alpha, also known as FcεR1α or FcER1a) which is likewise known as the IgE Fc receptor subunit alpha. FcεR1α is a type I transmembrane glycoprotein that is encoded by Fcer1a (Fc receptor, IgE, high affinity I, alpha polypeptide) which belongs to the Ig gene superfamily. As a single IgE-binding alpha subunit, FcεR1α complexes with signal transducing subunits including one beta subunit (FcεRIβ encoded by Ms4a2) and two disulfide-linked gamma subunits (FcεRIγ encoded by Fcer1g) to form the high-affinity receptor for IgE, Fc epsilon RI (FcεR1 or FcER1). FcεR1 is expressed on basophils and mast cells. Binding of cognate antigens (allergens) to FcεR1 with bound IgE antibodies leads to cellular activation and the release of mediators, including histamine and cytokines, that are responsible for allergic reactions. Tang et al. (2019) have reported that the MAR-1 antibody crossreacts with two other mouse Fc receptor chains, FcγRI (CD64) and FcγRIV (CD16-2), that are expressed by monocytes, macrophages, dendritic cells, or neutrophils. They suggested MAR-1 binding is specific for FcεRIa only on mast cells and basophils and that for other cell types reactive with the MAR-1 antibody, it may be appropriate to refer to these as MAR-1-positive (MAR-1+) cells.
Development References (6)
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Obata K, Mukai K, Tsujimura Y, et al. Basophils are essential initiators of a novel type of chronic allergic inflammation.. Blood. 2007; 110(3):913-20. (Clone-specific: Flow cytometry). View Reference
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Ota T, Aoki-Ota M, Duong BH, Nemazee D. Suppression of IgE B cells and IgE binding to Fc(epsilon)RI by gene therapy with single-chain anti-IgE.. J Immunol. 2009; 182(12):8110-7. (Clone-specific: Flow cytometry). View Reference
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Pellefigues C, Mehta P, Prout MS, et al. The Basoph8 Mice Enable an Unbiased Detection and a Conditional Depletion of Basophils.. Front Immunol. 2019; 10:2143. (Clone-specific: Flow cytometry). View Reference
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Perrigoue JG, Saenz SA, Siracusa MC, et al. MHC class II-dependent basophil-CD4+ T cell interactions promote T(H)2 cytokine-dependent immunity.. Nat Immunol. 2009; 10(7):697-705. (Clone-specific: Flow cytometry, In vivo exacerbation). View Reference
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Sokol CL1, Barton GM, Farr AG, Medzhitov R.. A mechanism for the initiation of allergen-induced T helper type 2 responses.. Nat Immunol. 2008; 9(3):310-318. (Clone-specific: Blocking, Flow cytometry, Functional assay, In vivo exacerbation). View Reference
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Tang XZ, Jung JB, Allen CDC. A case of mistaken identity: The MAR-1 antibody to mouse FcεRIα cross-reacts with FcγRI and FcγRIV.. J Allergy Clin Immunol. 2019; 143(4):1643-1646.e6. (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.