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Two color flow cytometric analysis of Blimp-1 expression in activated mouse splenocytes. Mouse splenic leucocytes were stimulated for 3 days with lipopolysaccharide and stained with FITC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553088/553087/561877). The cells were subsequently fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302; Left Panel) or BD Horizon PE-CF594 Rat Anti-Mouse Blimp-1 antibody (Cat. No. 564269; Right Panel). Two-color flow cytometric contour plots showing the correlated expression of Blimp-1 (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of intact activated splenocytes. Unstimulated splenocytes show a staining pattern similar to Ig Isotype stained cells (data not shown). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
BD Horizon™ PE-CF594 Rat Anti-Mouse Blimp-1
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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.
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- 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.
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The 5E7 monoclonal antibody specifically binds to mouse B lymphocyte-induced maturation protein 1 (Blimp-1). Blimp-1 is a 98 kDa zinc finger-containing protein that is encoded by the Prdm1 gene and functions as a transcriptional repressor. Blimp-1 is essential for regulating the terminal differentiation of activated B cells into plasma cells. In this process, Blimp-1 suppresses genes involved in B cell proliferation, such as Myc and Bcl6. It controls other genes, such as Xbp1, that are required for upregulating the plasma cell's protein synthesis machinery which is essential for antibody production and secretion. Blimp-1 is an important regulator in other cell types as well. It functions in the regulation of T cell activation and homeostasis and NK cell maturation. Blimp-1 is expressed in activated T cells and can function to inhibit IL-2 production while enhancing the differentiation and expression of several molecules produced by effector T cells.
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).
Development References (9)
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Crotty S, Johnston RJ, Schoenberger SP. Effectors and memories: Bcl-6 and Blimp-1 in T and B lymphocyte differentiation. Nat Immunol. 2010; 11(2):114-120. (Biology). View Reference
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Huang S. Blimp-1 is the murine homolog of the human transcriptional repressor PRDI-BF1. Cell. 1994; 78(1):9. (Biology). View Reference
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Johnston RJ, Poholek AC, DiToro D, et al. Bcl6 and Blimp-1 are reciprocal and antagonistic regulators of T follicular helper cell differentiation.. Science. 2009; 325(5943):1006-10. (Biology). View Reference
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Kallies A, Carotta S, Huntington ND, et al. A role for Blimp1 in the transcriptional network controlling natural killer cell maturation. Blood. 2011; 117(6):1869-1879. (Immunogen: Western blot). View Reference
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Kallies A, Hasbold J, Tarlinton DM, et al. Plasma cell ontogeny defined by quantitative changes in blimp-1 expression. J Exp Med. 2004; 200(8):967-977. (Biology). View Reference
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Kallies A, Hawkins ED, Belz GT, et al. Transcriptional repressor Blimp-1 is essential for T cell homeostasis and self-tolerance. Nat Immunol. 2006; 7(5):466-474. (Biology). View Reference
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Kim SJ, Gregersen PK, Diamond B. Regulation of dendritic cell activation by microRNA let-7c and BLIMP1. J Clin Invest. 2013; 123(2):823-833. (Biology). View Reference
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Martins GA, Cimmino L, Liao J, Magnusdottir E, Calame K. Blimp-1 directly represses Il2 and the Il2 activator Fos, attenuating T cell proliferation and survival. J Exp Med. 2008; 205(9):1959-1965. (Biology). View Reference
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Turner CA Jr, Mack DH, Davis MM. Blimp-1, a novel zinc finger-containing protein that can drive the maturation of B lymphocytes into immunoglobulin-secreting cells. Cell. 1994; 77(2):297-306. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.