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Alexa Fluor® 647 Rat Anti-Pax-5
Alexa Fluor® 647 Rat Anti-Pax-5

Multicolor flow cytometric analysis of Pax5 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 Alexa Fluor® 647 Rat Anti-Mouse Pax5 antibody (Cat. No. 562814) or Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690). The PBMC were counterstained with PE Mouse Anti-Human CD21 antibody (Cat. No. 555422/557327/561768). The mouse splenic leucocytes were counterstained with FITC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553088/553087/561877). 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 Pax5 (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 Pax5 (Right Plot) versus CD45R/B220 for events with the light-scatter characteristics of intact mouse splenocytes.

Multicolor flow cytometric analysis of Pax5 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 Alexa Fluor® 647 Rat Anti-Mouse Pax5 antibody (Cat. No. 562814) or Alexa Fluor® 647 Rat IgG2a, κ Isotype Control (Cat. No. 557690). The PBMC were counterstained with PE Mouse Anti-Human CD21 antibody (Cat. No. 555422/557327/561768). The mouse splenic leucocytes were counterstained with FITC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553088/553087/561877). 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 Pax5 (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 Pax5 (Right Plot) versus CD45R/B220 for events with the light-scatter characteristics of intact mouse splenocytes.

Product Details
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BD Pharmingen™
Pax5, PAX5, KLP, BSAP, EBB-1; B-cell-specific transcription factor
Mouse (QC Testing), Human (Tested in Development)
Rat IgG2a, κ
Recombinant Mouse Pax-5 protein containing aa 154-284
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2737811
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
562814 Rev. 1
Antibody Details
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1H9

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.

562814 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
562814 Rev.1
Citations & References
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Development References (8)

  1. 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
  2. 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
  3. 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
  4. 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
  5. 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
  6. 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
  7. 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
  8. 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
View All (8) View Less
562814 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.