Two-color flow cytometric analysis of CD23 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Anti-Mouse IgM[a] antibody (Cat. No. 553516) and either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse CD23 antibody (Cat. No. 563986; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of CD23 (or Ig Isotype control staining) versus IgM for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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Two-color flow cytometric analysis of CD23 expression on mouse splenocytes. Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with FITC Anti-Mouse IgM[a] antibody (Cat. No. 553516) and either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon™ PE-CF594 Rat Anti-Mouse CD23 antibody (Cat. No. 563986; Right Panel). Two-color flow cytometric dot plots show the correlated expression patterns of CD23 (or Ig Isotype control staining) versus IgM for gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
FcεRII; Fc-epsilon-RII; Fcer2a; Ly-42; Low-affinity IgE receptor; Fcer2
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
FcεR isolated from the mouse B hybridoma line O1.2B2
Flow cytometry (Routinely Tested)
0.2 mg/ml
14128
AB_2738524
Aqueous buffered solution containing BSA and ≤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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- CF™ is a trademark of Biotium, Inc.
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
PE-CF594 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562302
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563986 Rev. 1
Antibody Details
B3B4
The B3B4 monoclonal antibody specifically binds to CD23, the low affinity IgE Fc receptor (FcεRII) expressed on mature resting conventional B lymphocytes, but not on B-1 cells (CD5+ B cells) or T lymphocytes. It does not react with high-affinity IgE receptors, as demonstrated on mouse mast cell lines. The regulation of CD23 surface expression on activated B cells appears to be complex, depending upon the mode of activation and the presence of cytokines. IgE synthesis is negatively regulated by CD23, and CD23 expression is upregulated on splenocytes in the presence of IgE. CD23 is also upregulated on follicular dendritic cells in the lymph nodes of immunized mice, and a subset of splenic dendritic cells expresses CD23. The B3B4 antibody abrogates antigen-specific IgE-dependent modulation of immune responses in normal mice. This monoclonal antibody also blocks IgE binding and eosinophil infiltration in the lung of immunized mice. Different in vivo results have been obtained when using the intact B3B4 antibody or the F(ab')2 fragments. B3B4 mAb does not cross-react with rat or human IgE Fc Receptor.
This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg 610/20-nm filter).
563986 Rev. 1
Format Details
PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
563986 Rev.1
Citations & References
Development References (14)
-
Conrad DH, Waldschmidt TJ, Lee WT, et al. Effect of B cell stimulatory factor-1 (interleukin 4) on Fc epsilon and Fc gamma receptor expression on murine B lymphocytes and B cell lines. J Immunol. 1987; 139(7):2290-2296. (Clone-specific: Flow cytometry, Functional assay, Immunoaffinity chromatography, Immunoprecipitation, Radioimmunoassay). View Reference
-
Coyle AJ, Wagner K, Bertrand C, Tsuyuki S, Bews J, Heusser C. Central role of immunoglobulin (Ig) E in the induction of lung eosinophil infiltration and T helper 2 cell cytokine production: inhibition by a non-anaphylactogenic anti-IgE antibody. J Exp Med. 1996; 183(4):1303-1310. (Clone-specific: Blocking). View Reference
-
Dasic G, Juillard P, Graber P, et al. Critical role of CD23 in allergen-induced bronchoconstriction in a murine model of allergic asthma. Eur J Immunol. 1999; 29(9):2957-2967. (Clone-specific: Blocking, In vivo exacerbation). View Reference
-
Kisselgof AB, Oettgen HC. The expression of murine B cell CD23, in vivo, is regulated by its ligand, IgE. Int Immunol. 1998; 10(9):1377-1384. (Clone-specific: Flow cytometry). View Reference
-
Maeda K, Burton GF, Padgett DA, et al. Murine follicular dendritic cells and low affinity Fc receptors for IgE (Fc epsilon RII). J Immunol. 1992; 148(8):2340-2347. (Clone-specific: Electron microscopy, Immunohistochemistry). View Reference
-
Oshiba A, Hamelmann E, Haczku A, et al. Modulation of antigen-induced B and T cell responses by antigen-specific IgE antibodies. J Immunol. 1997; 159(8):4056-4063. (Clone-specific: Blocking). View Reference
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Pulendran B, Lingappa J, Kennedy MK, et al. Developmental pathways of dendritic cells in vivo: distinct function, phenotype, and localization of dendritic cell subsets in FLT3 ligand-treated mice. J Immunol. 1997; 159(5):2222-2231. (Clone-specific: Flow cytometry). View Reference
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Rabin E, Cong YZ, Wortis HH. Loss of CD23 is a consequence of B-cell activation. Implications for the analysis of B-cell lineages. Ann N Y Acad Sci. 1992; 651:130-142. (Biology: Flow cytometry). View Reference
-
Rao M, Lee WT, Conrad DH. Characterization of a monoclonal antibody directed against the murine B lymphocyte receptor for IgE. J Immunol. 1987; 138(6):1845-1851. (Immunogen: Blocking, Immunoprecipitation, Inhibition, Radioimmunoassay). View Reference
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Stief A, Texido G, Sansig G, et al. Mice deficient in CD23 reveal its modulatory role in IgE production but no role in T and B cell development. J Immunol. 1994; 152(7):3378-3390. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
-
Waldschmidt T, Snapp K, Foy T, Tygrett L, Carpenter C. B-cell subsets defined by the Fc epsilon R. Ann N Y Acad Sci. 1992; 651:84-98. (Clone-specific). View Reference
-
Waldschmidt TJ, Conrad DH, Lynch RG. Expression of B cell surface receptors. II. IL-4 can accelerate the developmental expression of the murine B cell IgE Fc receptor. J Immunol. 1989; 143(9):2820-2827. (Clone-specific: Flow cytometry, Immunoaffinity chromatography). View Reference
-
Waldschmidt TJ, Conrad DH, Lynch RG. The expression of B cell surface receptors. I. The ontogeny and distribution of the murine B cell IgE Fc receptor. J Immunol. 1988; 140(7):2148-2154. (Clone-specific: Flow cytometry). View Reference
-
Yu P, Kosco-Vilbois M, Richards M, Kohler G, Lamers MC. Negative feedback regulation of IgE synthesis by murine CD23. Nature. 1994; 369(6483):753-756. (Biology). View Reference
563986 Rev. 1
Please refer to Support Documents for Quality Certificates
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.
