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Multicolor flow cytometric analysis of Aiolos expression in human peripheral blood lymphocytes. Human whole blood was stained with APC Mouse Anti-Human CD19 antibody (Cat. No. 555415/561742) and FITC Mouse Anti-Human CD3 antibody (Cat. No. 555916). The cells were subsequently fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; Left Plots) or BD Horizon™ PE-CF594 Mouse Anti-Human Aiolos antibody (Cat. No. 567871/567872; Right Plots). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System. Upper Plots: Pseudocolor bivariate density plots showing the correlated expression of Aiolos (or Ig Isotype control staining) versus CD19 were derived from gated events with the forward and side-light scattering characteristics of intact lymphocytes. Bottom Plots: Pseudocolor bivariate density plots showing the correlated expression of Aiolos (or Ig Isotype control staining) versus CD3 were derived from gated events with the forward and side-light scattering characteristics of intact lymphocytes.
BD Horizon™ PE-CF594 Mouse Anti-Human Aiolos
Regulatory Status Legend
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
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
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- 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.
- 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).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
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Companion Products
The S50-895 monoclonal antibody specifically recognizes the transcription factor, Aiolos, which is encoded by IKZF3, IKAROS family zinc finger 3. Aiolos belongs to the Ikaros family of zinc-finger proteins that includes Ikaros and Helios. Aiolos can form homodimers or heterodimers with other Ikaros family members. Multiple Aiolos isoforms have been described. Aiolos is expressed in committed lymphoid progenitors and mature types of T and B lymphocytes, but is not expressed by hematopoietic stem cells. Aiolos plays an important role in the control of B lymphocyte differentiation and proliferation and is specifically required for the generation of long-lived, high affinity plasma cells in the bone marrow. Aiolos is involved in regulating BCL2 expression and controlling apoptosis in T-cells in an IL2-dependent manner. Aberrant expression of IKZF3 has been implicated with leukemogenesis and certain autoimmune diseases.
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).
Development References (8)
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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
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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
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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
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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
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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
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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
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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
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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
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.