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Multicolor flow cytometric analysis of FcεRIα expression on viable Mouse splenic leucocytes. C57BL/6 Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with FITC Hamster Anti-Mouse CD3e (Cat. No. 553062), FITC Rat Anti-Mouse CD45R/B220 (Cat. No. 553088), and APC Rat Anti-Mouse CD49b (Cat. No. 560628) antibodies and with either BD Horizon™ BV421 Hamster IgG1, κ Isotype Control (Cat. No. 562601; Left Plot) or BD OptiBuild™ BV421 Mouse Anti-FcεRIα antibody (Cat. No. 750852; Right Plot) at 0.25 µg/test. 7-AAD (7-Amino-Actinomycin D) Solution (Cat. No. 559925) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of FcεRIα (or Ig Isotype control staining) versus CD49b was derived from CD3-negative B220-negative gated events with the forward and side light-scatter characteristics of viable (7-AAD-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.
Flow cytometric analysis of FcεRIα expression in Mouse MC/9 cells. Cells from the Mouse MC/9 (Mast Cells, ATCC® CRL-8306™) cell line were stained with BD Horizon™ BV421 Hamster IgG1, κ Isotype Control (Cat. No. 562601; dashed line histogram) or BD OptiBuild™ BV421 Mouse Anti-FcεRIα antibody (Cat. No. 750852; 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.
BD OptiBuild™ BV421 Hamster Anti-Mouse FceR1a
BD OptiBuild™ BV421 Hamster Anti-Mouse FceR1a
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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® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
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.
- 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.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 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 http://regdocs.bd.com to access safety data sheets (SDS).
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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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.