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PE Rat Anti-Blimp-1
PE Rat Anti-Blimp-1

Flow Cytometric Analysis of Blimp-1 Expression

       Left Panel - 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). Fixed cells were stained with PE Rat Anti-Blimp-1 antibody (Cat. No. 564702).

       The two-color contour plot shows the coexpression of CD19 vs CD27 for gated events with the forward and side light-scatter characteristics of live lymphocytes. Histograms showing Blimp-1 expression for B cell subsets were derived from gated events as indicated. Flow cytometric analysis was performed with a BD LSRFortessa™ X-20 Flow Cytometer System.

       Right Panel - In Vitro-Differentiated Human Plasma Cells. CD19+CD27+ cells were sorted from PBMCs using the BD IMag™ Human B Lymphocyte Enrichment Set (Cat No 558007) followed by FACS sorting with a BD FACSAria™ III. Sorted cells were cultured (10 days) per Jourdan M, et al (2009) for differentiation of plasma cells. Cells were initially stained with BD Horizon™ Fixable Viability Stain 510 (FVS 510; Cat. No. 564406) for dead cell exclusion. Cells were surface stained with APC Anti-Human CD38 (Cat No. 555462) and BV421 Anti-Human CD138 (Cat No. 562935) antibodies. Cells were then fixed and permeabilized with the Transcription Factor Buffer Set and stained with either PE Rat IgG2a, κ Isotype Control (Cat No. 554689) or PE Rat Anti-Blimp-1 antibody.

       The two-color contour plot shows the coexpression of CD38 vs CD138 by live cells identified as FVS 510 negative. Plasma cells were identified as CD38+CD138+ cells and gated for the generation of the histograms showing Blimp-1 expression (solid line) versus Isotype Control staining (dashed line). Flow cytometric analysis was performed with a BD FACSCanto™ II Flow Cytometer System.

Flow Cytometric Analysis of Blimp-1 Expression

       Left Panel - 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). Fixed cells were stained with PE Rat Anti-Blimp-1 antibody (Cat. No. 564702).

       The two-color contour plot shows the coexpression of CD19 vs CD27 for gated events with the forward and side light-scatter characteristics of live lymphocytes. Histograms showing Blimp-1 expression for B cell subsets were derived from gated events as indicated. Flow cytometric analysis was performed with a BD LSRFortessa™ X-20 Flow Cytometer System.

       Right Panel - In Vitro-Differentiated Human Plasma Cells. CD19+CD27+ cells were sorted from PBMCs using the BD IMag™ Human B Lymphocyte Enrichment Set (Cat No 558007) followed by FACS sorting with a BD FACSAria™ III. Sorted cells were cultured (10 days) per Jourdan M, et al (2009) for differentiation of plasma cells. Cells were initially stained with BD Horizon™ Fixable Viability Stain 510 (FVS 510; Cat. No. 564406) for dead cell exclusion. Cells were surface stained with APC Anti-Human CD38 (Cat No. 555462) and BV421 Anti-Human CD138 (Cat No. 562935) antibodies. Cells were then fixed and permeabilized with the Transcription Factor Buffer Set and stained with either PE Rat IgG2a, κ Isotype Control (Cat No. 554689) or PE Rat Anti-Blimp-1 antibody.

       The two-color contour plot shows the coexpression of CD38 vs CD138 by live cells identified as FVS 510 negative. Plasma cells were identified as CD38+CD138+ cells and gated for the generation of the histograms showing Blimp-1 expression (solid line) versus Isotype Control staining (dashed line). Flow cytometric analysis was performed with a BD FACSCanto™ II Flow Cytometer System.

Product Details
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BD Pharmingen™
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
AB_2738901
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 R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

Clone 6D3 crossreacts with human and mouse Blimp-1. Customers could also consider fluorescent conjugates of clone 5E7 (Rat Anti-Mouse Blimp-1) for the staining and flow cytometric analysis of mouse cells that express Blimp-1.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
564702 Rev. 1
Antibody Details
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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.

564702 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
564702 Rev.1
Citations & References
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Development References (16)

  1. 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
  2. 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
  3. Huang S. Blimp-1 is the murine homolog of the human transcriptional repressor PRDI-BF1. Cell. 1994; 78(1):9. (Biology). View Reference
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
  9. Keller AD, Maniatis T. Identification and characterization of a novel repressor of beta-interferon gene expression. Genes Dev. 1991; 5(5):868-879. (Biology). View Reference
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
View All (16) View Less
564702 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.