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Multicolor flow cytometric analysis of Blimp-1 expression in human peripheral blood lymphocytes. Human PBMCs were stained in BD Horizon™ Brilliant Stain Buffer (Cat. No. 563794) with BV786 Mouse Anti-Human CD27 (Cat No. 563327) and BUV737 Mouse Anti-Human CD19 (Cat. No. 564303) antibodies and fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725). The cells were stained with BD Horizon™ PE-CF594 Rat Anti-Blimp-1 antibody (Cat. No. 565274). The two-color contour plot (Left Panel) showing the correlated expression of CD27 versus CD19 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Histograms (Right Panel) showing Blimp-1 expression for B cell subsets were derived from CD27 versus CD19 gated events as indicated. Flow cytometric analysis was performed with a BD LSRFortessa™ Flow Cytometer System.
BD Horizon™ PE-CF594 Rat Anti-Blimp-1
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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
For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794).
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.
- 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.
- 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.
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- 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.
- 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.
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- 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
The 6D3 monoclonal antibody specifically binds to mouse B lymphocyte-induced maturation protein 1 (Blimp-1) and crossreacts with human BLIMP-1. Blimp-1 is a 98 kDa zinc finger-containing protein that is encoded by the Prdm1 gene. Blimp-1 was first discovered as a transcriptional repressor of the IFN-β promoter. Blimp-1 is critical for primordial germ cells specification in the early embryo and induction of terminal cell differentiation in multiple somatic cell types. It is largely known as a master regulator of antibody-secreting plasma cells due to its function in the repression of PAX5, CIITA and cMYC genes. Blimp-1 also controls T cell homeostasis and effector differentiation through different mechanisms, including repression of IL-17 producing cells. Blimp-1 also plays roles in the maturation and activity of dendritic cells and natural killer 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 (12)
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Chan YH, Chiang MF, Tsai YC, et al. Absence of the transcriptional repressor Blimp-1 in hematopoietic lineages reveals its role in dendritic cell homeostatic development and function. J Immunol. 2009; 183(11):7039-7046. (Biology). View Reference
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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. (Biology). 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. (Immunogen: Western blot). 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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Ohinata Y, Payer B, O'Carroll D, et al. Blimp1 is a critical determinant of the germ cell lineage in mice. Nature. 2005; 436(7048):207-213. (Biology). View Reference
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Shaffer AL, Lin KI, Kuo TC, et al. Blimp-1 orchestrates plasma cell differentiation by extinguishing the mature B cell gene expression program. Immunity. 2002; 17(1):51-62. (Biology). View Reference
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Smith MA, Maurin M, Cho HI, et al. PRDM1/Blimp-1 controls effector cytokine production in human NK cells.. J Immunol. 2010; 185(10):6058-67. (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.