Flow cytometric analysis of Aiolos expression by mouse lymphoid cells. Left Panel - Mouse thymocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). Thymocytes were then stained with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon PE-CF594 Mouse Anti Mouse Aiolos antibody (Cat. No. 565267; solid line histogram). Fluorescence histograms showing the expression of Aiolos (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of intact thymocytes. Middle and Right Panels - Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826). The cells were fixed and permeabilized, using the BD Pharmingen™ Transcription Factor Buffer Set and further stained with BD Horizon PE-CF594 Mouse IgG1, κ Isotype Control (Middle Panel), or BD Horizon PE-CF594 Mouse Anti-Mouse Aiolos antibody (Right Panel) as indicated. Two-color flow cytometric contour plots showing the correlated expression of Aiolos (or Ig Isotype control staining) versus CD3e were derived from 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.
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Flow cytometric analysis of Aiolos expression by mouse lymphoid cells. Left Panel - Mouse thymocytes were fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574/562725). Thymocytes were then stained with either BD Horizon™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon PE-CF594 Mouse Anti Mouse Aiolos antibody (Cat. No. 565267; solid line histogram). Fluorescence histograms showing the expression of Aiolos (or Ig Isotype control staining) were derived from gated events with the forward and side-light scattering characteristics of intact thymocytes. Middle and Right Panels - Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142) and then stained with APC Hamster Anti-Mouse CD3e antibody (Cat. No. 553066/561826). The cells were fixed and permeabilized, using the BD Pharmingen™ Transcription Factor Buffer Set and further stained with BD Horizon PE-CF594 Mouse IgG1, κ Isotype Control (Middle Panel), or BD Horizon PE-CF594 Mouse Anti-Mouse Aiolos antibody (Right Panel) as indicated. Two-color flow cytometric contour plots showing the correlated expression of Aiolos (or Ig Isotype control staining) versus CD3e were derived from 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.
Product Details
BD Horizon™
Ikzf3; IKAROS family zinc finger 3; Zfpn1a3; Znfn1a3
Mouse (QC Testing)
Mouse IgG1, κ
Human Aiolos Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2739142
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 BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- 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.
- 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.
- CF™ is a trademark of Biotium, Inc.
- 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.
- Texas Red is a registered 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
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.1 mg
Cat No.
553141
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Transcription Factor Buffer Set RUO
Size
100 Tests
Cat No.
562574
PE-CF594 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
562292
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565267 Rev. 1
Antibody Details
S48-791
The S48-791 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. Aiolos is expressed in committed lymphoid progenitors, thymocytes, and mature types of T and B lymphocytes and NK cells. It 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. Aiolos is also expressed by Th17 cells and reportedly promotes their differentiation by downregulating IL-2 expression.
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).
565267 Rev. 1
Format Details
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.
PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
565267 Rev.1
Citations & References
Development References (7)
-
Cariappa A, Tang M, Parng C, et al. The follicular versus marginal zone B lymphocyte cell fate decision is regulated by Aiolos, Btk, and CD21. Immunity. 2001; 14(5):603-615. (Biology). View Reference
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Cortes M, Georgopoulos K. Aiolos is required for the generation of high affinity bone marrow plasma cells responsible for long-term immunit. J Exp Med. 204; 199(2):209-219. (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
-
Quintana FJ, Jin H, Burns EJ, et al. Aiolos promotes TH17 differentiation by directly silencing Il2 expression. Nat Immunol. 2012; 13(8):770-777. (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
-
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
565267 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.
