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PE-CF594 Mouse Anti-Human Aiolos
PE-CF594 Mouse Anti-Human Aiolos
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.
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.
Product Details
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BD Horizon™
AIOLOS; AIO; IKZF3; ZNFN1A3
Human (QC Testing)
Mouse IgG1, κ
Human Aiolos Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
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 the dye under optimum conditions and unconjugated antibody and free dye were removed.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. 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.
  12. 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.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567872 Rev. 1
Antibody Details
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S50-895

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).

567872 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
567872 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
567872 Rev. 1

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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.