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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™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human Ikaros antibody (Cat. No. 564865; 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.
BD Horizon™ BV421 Mouse Anti-Human Ikaros
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (5)
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Georgopoulos K. Haematopoietic cell-fate decisions, chromatin regulation and ikaros. Nat Rev Immunol. 2002; 2(3):162-174. (Biology). View Reference
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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
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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
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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
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Yoshida T, Landhuis E, Dose M, et al. Transcriptional regulation of the Ikzf1 locus. Blood. 2013; 122(18):3149-3159. (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.