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.
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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.
Product Details
BD Horizon™
Prdm1; PR domain zinc finger protein 1; Blimp1; PRDI-BF1; ZNFPR1A1
Mouse (QC Testing)
Rat IgG2a, κ
Mouse Blimp-1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2738719
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.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- 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.
- 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.
- CF™ is a trademark of Biotium, Inc.
- 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
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
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
PE-CF594 Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562302
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FITC Rat Anti-Mouse CD45R/B220 RUO
Size
0.5 mg
Cat No.
553088
564269 Rev. 1
Antibody Details
5E7
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).
564269 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
564269 Rev.1
Citations & References
Development References (9)
-
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
-
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
-
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
564269 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.
