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PE Rat Anti-Mouse Spi-B
PE Rat Anti-Mouse Spi-B
Flow cytometric analysis of Spi-B expression in mouse leucocytes. C57BL/6 mouse splenocytes (Left Panel) and bone marrow cells (Right Panel) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with BD Horizon™ BV711 Rat Anti-Mouse CD45R/B220 antibody (Cat No. 563892). After washing, the cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725) and stained with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plots) or PE Rat Anti-Mouse Spi-B antibody (Cat. No. 565834; Right Plots). Two-color flow cytometric contour plots showing the correlated expression of Spi-B (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Flow cytometric analysis of Spi-B expression in mouse leucocytes. C57BL/6 mouse splenocytes (Left Panel) and bone marrow cells (Right Panel) were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and stained with BD Horizon™ BV711 Rat Anti-Mouse CD45R/B220 antibody (Cat No. 563892). After washing, the cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574/562725) and stained with either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plots) or PE Rat Anti-Mouse Spi-B antibody (Cat. No. 565834; Right Plots). Two-color flow cytometric contour plots showing the correlated expression of Spi-B (or Ig Isotype control staining) versus CD45R/B220 were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
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BD Pharmingen™
SPI-B; Spib; Spi-1/PU.1 related; Ets transcription factor Spi-B
Mouse (QC Testing)
Rat IgG2a, κ
Mouse Spi-B Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739374
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.

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. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
565834 Rev. 1
Antibody Details
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U14-267

The U14-267 monoclonal antibody specifically recognizes mouse Spi-B, that is encoded by Spib (Spi-B transcription factor (Spi-1/PU.1 related)). Spi-B belongs to the Ets family of transcription factors. It is expressed predominantly in B cells, T-cell progenitors, and plasmacytoid dendritic cells. Spi-B plays crucial roles in the development of T cells and plasmacytoid dendritic cells, and in the terminal differentiation of B cells.  Although Spi-B deficient mice have normal numbers of B cells, they exhibit defects in germinal center formation and maintenance. In contrast, overexpression of Spi-B blocks plasma cell differentiation due to its capacity to repress Blimp-1 and Xbp-1 expression.

565834 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.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
565834 Rev.1
Citations & References
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Development References (5)

  1. Bartel FO, Higuchi T, Spyropoulos DD. Mouse models in the study of the Ets family of transcription factors.. Oncogene. 2000; 19(55):6443-54. (Biology). View Reference
  2. Lefebvre JM, Haks MC, Carleton MO, et al. Enforced expression of Spi-B reverses T lineage commitment and blocks beta-selection.. J Immunol. 2005; 174(10):6184-94. (Biology). View Reference
  3. Schmidlin H, Diehl SA, Nagasawa M, et al. Spi-B inhibits human plasma cell differentiation by repressing BLIMP1 and XBP-1 expression.. Blood. 2008; 112(5):1804-12. (Biology). View Reference
  4. Schotte R, Nagasawa M, Weijer K, Spits H, Blom B. The ETS transcription factor Spi-B is required for human plasmacytoid dendritic cell development.. J Exp Med. 2004; 200(11):1503-9. (Biology). View Reference
  5. Su GH, Chen HM, Muthusamy N, et al. Defective B cell receptor-mediated responses in mice lacking the Ets protein, Spi-B.. EMBO J. 1997; 16(23):7118-29. (Biology). View Reference
View All (5) View Less
565834 Rev. 1

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