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Multicolor flow cytometric analysis of Pax-5 expression in human and mouse leucocytes. Human peripheral blood mononuclear cells (PBMC) and BALB/c mouse splenic leucocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with either BD Horizon™ PE-CF594 Rat Anti-Mouse Pax-5 antibody (Cat. No. 562815) or BD Horizon™ PE-CF594 Rat IgG2a, κ Isotype Control (Cat. No. 562302). The PBMC were counterstained with APC Mouse Anti-Human CD21 antibody (Cat. No. 561767/561357/559867). The mouse splenic leucocytes were counterstained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092/561880). Flow cytometric analysis was performed using a BD LSR™ II Flow Cytometer System. Panel A. Human Peripheral Blood Lymphocytes: The two-color flow cytometric dot plots show the correlated expression patterns of Ig Isotype control staining (Left Plot) and Pax-5 (Right Plot) versus CD21 for events with the forward and side light-scatter characteristics of intact human peripheral blood lymphocytes. Panel B. Mouse Splenocytes: The dot plots show the coexpression patterns of Ig Isotype control staining (Left Plot) or Pax-5 (Right Plot) versus CD45R/B220 for events with the light-scatter characteristics of intact mouse splenocytes.
BD Horizon™ PE-CF594 Rat Anti-Pax-5
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- CF™ is a trademark of Biotium, Inc.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- 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.
- 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.
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The 1H9 monoclonal antibody clone specifically binds to human and mouse Paired box protein Pax-5. Pax-5 is a ~50 kDa protein that is also known as B-cell-specific transcription factor and B cell specific activator protein (BSAP). Pax-5 is a member of the paired box (Pax) family of transcription factors. It is expressed in pro-B, pre-B and mature B cells. Through the Groucho family of co-repressors, Pax-5 likely functions as a transcriptional repressor of non-B-lymphoid genes during the B cell commitment process. In the early stages of B cell development, Pax-5 influences the expression of several B-cell-specific genes, such as CD19 and CD20 and maintains B cell identity. Pax-5 suppression is involved in the upregulation of Blimp-1 leading to the development of Pax-5-negative plasma cells. Pax-5 mRNA is transiently detected in the mesencephalon and spinal cord during embryogenesis. Expression then shifts to the fetal liver and correlates with the onset of B lymphopoiesis. Altered forms and expression patterns of Pax-5 have been associated with some lymphoid and nonlymphoid cancers.
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).
Development References (8)
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Adams B, Dorfler P, Aguzzi A, et al. Pax-5 encodes the transcription factor BSAP and is expressed in B lymphocytes, the developing CNS, and adult testis. Genes Dev. 1992; 6(9):1589-1607. (Biology). View Reference
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Foss HD, Reusch R, Demel G, et al. Frequent expression of the B-cell-specific activator protein in Reed-Sternberg cells of classical Hodgkin's disease provides further evidence for its B-cell origin. Blood. 1999; 94(9):3108-3113. (Biology). View Reference
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Hertel CB, Zhou XG, Hamilton-Dutoit SJ, Junker S. Loss of B cell identity correlates with loss of B cell-specific transcription factors in Hodgkin/Reed-Sternberg cells of classical Hodgkin lymphoma. Oncogene. 2002; 21(32):4908-4920. (Biology). View Reference
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Kallies A, Hasbold J, Fairfax K, et al. Initiation of plasma-cell differentiation is independent of the transcription factor Blimp-1. Immunity. 2007; 26(5):555-566. (Immunogen: Western blot). View Reference
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Klein U, Tu Y, Stolovitzky GA, et al. Transcriptional analysis of the B cell germinal center reaction. Proc Natl Acad Sci U S A. 2003; 100(5):2639-2644. (Biology). View Reference
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McManus S, Ebert A, Salvagiotto G, et al. The transcription factor Pax5 regulates its target genes by recruiting chromatin-modifying proteins in committed B cells. EMBO J. 2011; 30(12):2388-2404. (Clone-specific: Western blot). View Reference
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O'Brien P, Morin P, Jr., Ouellette RJ, Robichaud GA. The Pax-5 gene: a pluripotent regulator of B-cell differentiation and cancer disease. Cancer Res. 2011; 71(24):7345-7350. (Biology). View Reference
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Zwollo P, Arrieta H, Ede K, Molinder K, Desiderio S, Pollock R. The Pax-5 gene is alternatively spliced during B-cell development. J Biol Chem. 1997; 272(15):10160-10168. (Biology). View Reference
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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.
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