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Purified Rat Anti-Blimp-1
Purified Rat Anti-Blimp-1

Analyses of Blimp-1 Expression

     Panel 1.  Western blot analysis of Blimp-1 expression in human myeloma cell lines. Lysates from RPMI-8226 (ATCC CLL-155; Lane 1) and U266 (ATCC TIB-196; Lane 2) cells were electrophoresed, blotted and probed with Purified Rat Anti-Blimp-1 antibody (Cat. No. 564703) followed by HRP Goat Anti-Rat Ig (Cat. No. 554017), and ECL detection reagents. The antibody detects Blimp-1 isoforms of 88-92 kDa in both cell lysates and 76-78 kDa) in the U266 cell lysate. Note: an unspecific band (~250 kDa) may be observed upon overexposure of the membranes.

    Panel 2. Flow cytometric analysis of Blimp-1 expression in a human myeloma cell line. RPMI 8226 cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725) and stained with either Purified Rat IgG2a, κ (Cat. No. 553927; dashed line histogram) or Purified Rat Anti-Blimp-1 antibody (solid line histogram) followed by staining with PE Goat Anti-Rat Ig (Cat. No. 560767). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.

    Panel 3. Immunohistochemical staining of human Blimp-1 in human tonsils. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded tonsil sections were stained with either Purified Rat IgG2a κ Isotype Control (Left Image) or Purified Rat Anti-Human Blimp-1 antibody (Right Image). A three-step staining procedure that employs Biotin Goat Anti-Rat Immunoglobulin (Cat. No. 559286), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the Blimp-1 specific antibody primarily stained the nuclei of some Germinal Center cells. Original magnification: 40×.

Analyses of Blimp-1 Expression

     Panel 1.  Western blot analysis of Blimp-1 expression in human myeloma cell lines. Lysates from RPMI-8226 (ATCC CLL-155; Lane 1) and U266 (ATCC TIB-196; Lane 2) cells were electrophoresed, blotted and probed with Purified Rat Anti-Blimp-1 antibody (Cat. No. 564703) followed by HRP Goat Anti-Rat Ig (Cat. No. 554017), and ECL detection reagents. The antibody detects Blimp-1 isoforms of 88-92 kDa in both cell lysates and 76-78 kDa) in the U266 cell lysate. Note: an unspecific band (~250 kDa) may be observed upon overexposure of the membranes.

    Panel 2. Flow cytometric analysis of Blimp-1 expression in a human myeloma cell line. RPMI 8226 cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725) and stained with either Purified Rat IgG2a, κ (Cat. No. 553927; dashed line histogram) or Purified Rat Anti-Blimp-1 antibody (solid line histogram) followed by staining with PE Goat Anti-Rat Ig (Cat. No. 560767). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of intact cells. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.

    Panel 3. Immunohistochemical staining of human Blimp-1 in human tonsils. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), the formalin-fixed paraffin-embedded tonsil sections were stained with either Purified Rat IgG2a κ Isotype Control (Left Image) or Purified Rat Anti-Human Blimp-1 antibody (Right Image). A three-step staining procedure that employs Biotin Goat Anti-Rat Immunoglobulin (Cat. No. 559286), Streptavidin-Horseradish Peroxidase (HRP) (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) was used to develop the primary staining reagents. As shown in the Right Image, the Blimp-1 specific antibody primarily stained the nuclei of some Germinal Center cells. Original magnification: 40×.

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), Immunohistochemistry-paraffin, Western blot (Tested During Development)
0.5 mg/ml
AB_2738902
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Note: Staining with PE Rat Anti-Blimp-1 antibody (Cat. No. 564702) is recommended for the flow cytometric analysis of Blimp-1 expression in human peripheral blood plasma cells.

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. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
564703 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.

564703 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
564703 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
564703 Rev. 1

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