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Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α

BD Pharmingen™ Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α

Clone MAR-1 (also known as MAR1)

(RUO)
Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α
Flow cytometric analysis of FcεR1α expression on MC/9 cells. Cells from the mouse MC/9 (Mast Cells, ATCC CRL-8306) cell line were stained with Alexa Fluor™ 647 Hamster IgG1, κ Isotype Control (Cat. No. 563612; dashed line histogram) or Alexa Fluor™ 647 Hamster Anti-Mouse Anti-FcεR1α antibody (Cat. No. 567804; solid line histogram) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing FcεR1α expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α
Multiparameter flow cytometric analysis of FcεR1α expression on Mouse splenic leucocytes. 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 BD Horizon™ BV421 Hamster Anti-Mouse CD3e (Cat. No. 562600), BD Horizon™ BV421 Rat Anti-Mouse CD45R/B220 (Cat. No. 562922), FITC Rat Anti-Mouse CD49b (Cat. No. 561067) antibodies and with either Alexa Fluor™ 647 Hamster IgG1, κ Isotype Control (Cat. No. 563612; Left Plot) or Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α antibody (Cat. No. 567804; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of FcεR1α (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 (DAPI-negative) leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of FcεR1α expression on MC/9 cells. Cells from the mouse MC/9 (Mast Cells, ATCC CRL-8306) cell line were stained with Alexa Fluor™ 647 Hamster IgG1, κ Isotype Control (Cat. No. 563612; dashed line histogram) or Alexa Fluor™ 647 Hamster Anti-Mouse Anti-FcεR1α antibody (Cat. No. 567804; solid line histogram) at 0.5 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing FcεR1α expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multiparameter flow cytometric analysis of FcεR1α expression on Mouse splenic leucocytes. 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 BD Horizon™ BV421 Hamster Anti-Mouse CD3e (Cat. No. 562600), BD Horizon™ BV421 Rat Anti-Mouse CD45R/B220 (Cat. No. 562922), FITC Rat Anti-Mouse CD49b (Cat. No. 561067) antibodies and with either Alexa Fluor™ 647 Hamster IgG1, κ Isotype Control (Cat. No. 563612; Left Plot) or Alexa Fluor™ 647 Hamster Anti-Mouse FcεR1α antibody (Cat. No. 567804; Right Plot) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of FcεR1α (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 (DAPI-negative) leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Pharmingen™
FceR1 alpha; Fcer1a; FcεRIα; High affinity IgE Receptor; MAR1
Mouse (QC Testing)
Armenian Hamster IgG
Flow cytometry (Routinely Tested)
0.2 mg/ml
14125
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed.

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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  10. For U.S. patents that may apply, see bd.com/patents.
567804 Rev. 2
Antibody Details
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MAR-1

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.

567804 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
567804 Rev.2
Citations & References
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View product citations for antibody "567804" on CiteAb

Development References (6)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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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567804 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.