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PE Mouse Anti-Human Ikaros
PE 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 PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Panel) or PE Mouse Anti-Human Ikaros antibody (Cat. No. 564476; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of CD3 versus Ikaros (or Ig Isotype control staining) were generated for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ 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 PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Panel) or PE Mouse Anti-Human Ikaros antibody (Cat. No. 564476; Right Panel). Two-color flow cytometric contour plots showing the correlated expression patterns of CD3 versus Ikaros (or Ig Isotype control staining) were generated for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ 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_2738824
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 R-PE under optimum conditions, and unconjugated antibody and free PE 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
564476 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.

564476 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE 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 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
564476 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
564476 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.