Multicolor flow cytometric analysis of Aiolos expression in human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with either APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811; Left Panels) or BD Horizon™ V450 Mouse Anti-CD19 antibody (Cat. No. 560353; Right Panels). The cells were subsequently fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Lower Panels) or PE Mouse Anti-Human Aiolos antibody (Cat. No. 564811; Upper Panels). Two-color flow cytometric contour plots showing the correlated expression of CD3 or CD19 versus Aiolos (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
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Multicolor flow cytometric analysis of Aiolos expression in human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leucocytes were washed and then stained with either APC Mouse Anti-Human CD3 antibody (Cat. No. 555335/561810/561811; Left Panels) or BD Horizon™ V450 Mouse Anti-CD19 antibody (Cat. No. 560353; Right Panels). The cells were subsequently fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725) and stained with either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Lower Panels) or PE Mouse Anti-Human Aiolos antibody (Cat. No. 564811; Upper Panels). Two-color flow cytometric contour plots showing the correlated expression of CD3 or CD19 versus Aiolos (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.
Product Details
BD Pharmingen™
AIOLOS; AIO; IKZF3; ZNFN1A3
Human (QC Testing)
Mouse IgG1, κ
Human Aiolos Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
5 µl
AB_2738966
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
- 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).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
PE Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
554680
Transcription Factor Buffer Set RUO
Size
25 Tests
Cat No.
562725
564811 Rev. 1
Antibody Details
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.
564811 Rev. 1
Format Details
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.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
564811 Rev.1
Citations & References
View product citations for antibody "564811" on CiteAb
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
564811 Rev. 1
Please refer to Support Documents for Quality Certificates
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.