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PE-CF594 Mouse Anti-Human Ikaros
PE-CF594 Mouse Anti-Human Ikaros
Two-color flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Panel) or BD Horizon PE-CF594 Mouse Anti-Human Ikaros antibody (Cat. No. 564866; 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.
Two-color flow cytometric analysis of Ikaros expression on human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811) and either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Panel) or BD Horizon PE-CF594 Mouse Anti-Human Ikaros antibody (Cat. No. 564866; 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 Horizon™
IKZF1; LYF1; hIk-1; IK1; ZNFN1A1
Human (QC Testing)
Mouse IgG1, κ
Human Ikaros Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738989
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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

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. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  6. 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.
  7. 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.
  8. CF™ is a trademark of Biotium, Inc.
  9. 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.
  10. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  11. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  12. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564866 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.

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).

564866 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, 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 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
564866 Rev.1
Citations & References
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Development References (5)

  1. Georgopoulos K. Haematopoietic cell-fate decisions, chromatin regulation and ikaros. Nat Rev Immunol. 2002; 2(3):162-174. (Biology). View Reference
  2. Molnar A, Wu P, Largespada DA, et al. The Ikaros gene encodes a family of lymphocyte-restricted zinc finger DNA binding proteins, highly conserved in human and mouse. J Immunol. 1996; 156(2):585-592. (Biology). View Reference
  3. Nietfeld W, Meyerhans A. Cloning and sequencing of hIk-1, a cDNA encoding a human homologue of mouse Ikaros/LyF-1. Immunol Lett. 1996; 49(1-2):139-141. (Biology). View Reference
  4. Westman BJ, Mackay JP, Gell D. Ikaros: a key regulator of haematopoiesis. Int J Biochem Cell Biol. 2002; 34(10):1304-1307. (Biology). View Reference
  5. Yoshida T, Landhuis E, Dose M, et al. Transcriptional regulation of the Ikzf1 locus. Blood. 2013; 122(18):3149-3159. (Biology). View Reference
View All (5) View Less
564866 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.