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      Alexa Fluor® 647 Mouse Anti-Human Ikaros
      Alexa Fluor® 647 Mouse Anti-Human Ikaros
      Two-color flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809). The cells 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 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human Ikaros antibody (Cat. No. 564868; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of Ikaros (or Ig Isotype control staining) versus CD3 were generated for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Alexa Fluor® 647 Mouse Anti-Human Ikaros
      Two-color flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809). The cells 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 Mouse IgG1, κ Isotype Control (Cat. No. 557732; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human Ikaros antibody (Cat. No. 564868; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of Ikaros (or Ig Isotype control staining) versus CD3 were generated for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
      Product Details
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      BD Pharmingen™
      IKZF1; LYF1; hIk-1; IK1; ZNFN1A1
      Human (QC Testing), Mouse (Lack of Reactivity Confirmed in Development)
      Mouse IgG1, κ
      Human Ikaros Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl
      AB_2738991
      Aqueous buffered solution containing BSA and ≤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.
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      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. An isotype control should be used at the same concentration as the antibody of interest.
      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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      5. 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.
      6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      564868 Rev. 1
      Antibody Details
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      R32-1149

      The R32-1149 monoclonal antibody specifically binds to Ikaros, which is encoded by IKZF1 (IKAROS family zinc finger 1). Ikaros is also known as Lymphoid transcription factor LyF-1 (LYF1) and ZNFN1A1 [Zinc finger protein, subfamily 1A, 1 (Ikaros)]. Ikaros belongs to the IKAROS family of zinc-finger transcription factors. This DNA-binding transcription factor forms homodimers and heterodimers with other IKAROS family members. It is involved in the development of lymphoid tissues and remains abundantly expressed in the thymus and by cells within peripheral lymphoid tissues. Ikaros functions in the maturation, differentiation, and homeostasis of various hematopoietic cells including T lymphocytes, B cells, NK cells, and neutrophils. Ikaros regulates the expression of a number of genes including various genes expressed during early stages of B- and T-cell development. Several alternatively-spliced human Ikaros isoforms have been described. Abnormal expression of different Ikaros isoforms has been associated with hematologic malignancies. Flow cytometric analysis of 293F cells transfected with members of the IKAROS family has demonstrated that clone R32-1149 is negative on Helios, Aiolos, and Eos transfectants.

      564868 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
      564868 Rev.1
      Citations & References
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      Development References (8)

      1. Billot K, Parizot C, Arrouss I, et al. Differential aiolos expression in human hematopoietic subpopulations. Leuk Res. 2010; 34(3):289-293. (Biology). View Reference
      2. Billot K, Soeur J, Chereau F, et al. Deregulation of Aiolos expression in chronic lymphocytic leukemia is associated with epigenetic modifications. Blood. 2011; 117(6):1917-1927. (Biology). View Reference
      3. Liippo J, Nera KP, Veistinen E, et al. Both normal and leukemic B lymphocytes express multiple isoforms of the human Aiolos gene. Eur J Immunol. 2001; 31(12):3469-3474. (Biology). View Reference
      4. Morgan B, Sun L, Avitahl N, et al. Aiolos, a lymphoid restricted transcription factor that interacts with Ikaros to regulate lymphocyte differentiation. EMBO J. 1997; 16(8):2004-2013. (Biology). View Reference
      5. Rebollo A, Ayllon V, Fleischer A, Martinez CA, Zaballos A. The association of Aiolos transcription factor and Bcl-xL is involved in the control of apoptosis. J Immunol. 2001; 167(11):6366-6373. (Biology). View Reference
      6. Romero F, Martinez AC, Camonis J, Rebollo A. Aiolos transcription factor controls cell death in T cells by regulating Bcl-2 expression and its cellular localization. EMBO J. 1999; 18(12):3419-3430. (Biology). View Reference
      7. Schmitt C, Tonnelle C, Dalloul A, Chabannon C, Debre P, Rebollo A. Aiolos and Ikaros: regulators of lymphocyte development, homeostasis and lymphoproliferation. Apoptosis. 2002; 7(3):277-284. (Biology). View Reference
      8. Wang JH, Avitahl N, Cariappa A, et al. Aiolos regulates B cell activation and maturation to effector state. Immunity. 1998; 9(4):543-553. (Biology). View Reference
      View All (8) View Less
      564868 Rev. 1

      Please refer to Support Documents for Quality Certificates


      Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


      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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