Multicolor flow cytometric analysis of Blimp-1 expression in Human Peripheral Blood Lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stained in BD Horizon™ Brilliant Stain Buffer (Cat. No. 566349) with BD Horizon™ BUV737 Mouse Anti-Human CD27 (Cat No. 612829) and BD Horizon™ BUV395 Mouse Anti-Human CD19 (Cat. No. 563549) antibodies and washed. The cells were then fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725). Fixed cells were stained with BD Horizon™ R718 Rat Anti-Blimp-1 antibody (Cat. No. 567764) at 1 µg/test. The bivariate pseudocolor density plot (Left Panel) showing the correlated expression of CD19 versus CD27 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Histograms (Right Panel) showing Blimp-1 expression for the three B cell subsets defined by the correlated expression of CD19 versus CD27 were derived from gated events as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Multicolor flow cytometric analysis of Blimp-1 expression in Human Peripheral Blood Lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stained in BD Horizon™ Brilliant Stain Buffer (Cat. No. 566349) with BD Horizon™ BUV737 Mouse Anti-Human CD27 (Cat No. 612829) and BD Horizon™ BUV395 Mouse Anti-Human CD19 (Cat. No. 563549) antibodies and washed. The cells were then fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725). Fixed cells were stained with BD Horizon™ R718 Rat Anti-Blimp-1 antibody (Cat. No. 567764) at 1 µg/test. The bivariate pseudocolor density plot (Left Panel) showing the correlated expression of CD19 versus CD27 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Histograms (Right Panel) showing Blimp-1 expression for the three B cell subsets defined by the correlated expression of CD19 versus CD27 were derived from gated events as indicated. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
BLIMP-1; BLIMP1; PRDM1; PR domain containing 1, with ZNF domain; PRDI-BF1
Human (QC Testing), Mouse (Tested in Development)
Rat IgG2a, κ
Mouse Blimp-1 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
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 to the dye under optimum conditions and unreacted dye was removed.
Recommended Assay Procedures
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Brilliant Stain Buffer RUO
Size
1000 Tests
Cat No.
566349
BUV737 Mouse Anti-Human CD27 RUO
Size
100 Tests
Cat No.
612829
BUV395 Mouse Anti-Human CD19 RUO
Size
100 Tests
Cat No.
563549
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
567764 Rev. 1
Antibody Details
6D3
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.
567764 Rev. 1
Format Details
R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
R718
Red 627-640 nm
695 nm
718 nm
567764 Rev.1
Citations & References
View product citations for antibody "567764" on CiteAb
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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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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Jourdan M, Caraux A, De Vos J, et al. An in vitro model of differentiation of memory B cells intoplasmablasts and plasma cells including detailed phenotypic and molecularcharacterizatio. Blood. 2009; 114(25):5173-5181. (Methodology: Cell differentiation). 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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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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Roquilly A, McWilliam HEG, Jacqueline C, et al. Local Modulation of Antigen-Presenting Cell Development after Resolution of Pneumonia Induces Long-Term Susceptibility to Secondary Infections.. Immunity. 2017; 47(1):135-147.e5. (Clone-specific: Flow cytometry). View Reference
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Salehi S, Bankoti R, Benevides L, et al. B lymphocyte-induced maturation protein-1 contributes to intestinal mucosa homeostasis by limiting the number of IL-17-producing CD4+ T cells. J Immunol. 2012; 189(12):5682-5693. (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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Zumaquero E, Stone SL, Scharer CD, et al. IFNγ induces epigenetic programming of human T-bet(hi) B cells and promotes TLR7/8 and IL-21 induced differentiation. eLife. 2019; 8:e41641. (Clone-specific: Flow cytometry). View Reference
567764 Rev. 1
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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.