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PE-Cy™7 Rat Anti-Mouse CD23
PE-Cy™7 Rat Anti-Mouse CD23
Two-color flow cytometric analysis of CD23 expression on mouse splenocytes. Splenic leucocytes from a BALB/c mouse 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 Mouse Anti-Mouse IgM[a] antibody (Cat. No. 553516) and either PE-Cy™7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Panel) or PE-Cy™7 Rat Anti-Mouse CD23 antibody (Cat. No. 562825; 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.
Two-color flow cytometric analysis of CD23 expression on mouse splenocytes. Splenic leucocytes from a BALB/c mouse 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 Mouse Anti-Mouse IgM[a] antibody (Cat. No. 553516) and either PE-Cy™7 Rat IgG2a, κ Isotype Control (Cat. No. 552784; Left Panel) or PE-Cy™7 Rat Anti-Mouse CD23 antibody (Cat. No. 562825; 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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BD Pharmingen™
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_2737820
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 with PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. Cy is a trademark of Amersham Biosciences Limited. This conjugated product is sold under license to the following patents: US Patent Nos. 5,486,616; 5,569,587; 5,569,766; 5,627,027.
  5. 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.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  9. This product is subject to proprietary rights of Amersham Biosciences Corp. and Carnegie Mellon University and made and sold under license from Amersham Biosciences Corp. This product is licensed for sale only for research. It is not licensed for any other use. If you require a commercial license to use this product and do not have one return this material, unopened to BD Biosciences, 10975 Torreyana Rd, San Diego, CA 92121 and any money paid for the material will be refunded.
  10. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
562825 Rev. 1
抗体の詳細
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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.

            

562825 Rev. 1
フォーマットの詳細
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PE-Cy7
PE-Cy7 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, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-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 Red (627–640-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.
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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
562825 Rev.1
引用&参考文献
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Development References (14)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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. (Biology: Electron microscopy, Immunohistochemistry). View Reference
  6. 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
  7. 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
  8. 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. (Clone-specific: Flow cytometry). View Reference
  9. 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
  10. 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
  11. 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. (Biology). View Reference
  12. 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). View Reference
  13. 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, Immunoaffinity chromatography). View Reference
  14. 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
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562825 Rev. 1

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