Flow cytometric analysis of TOX expression in human MOLT-4 cells. Cells from the human MOLT-4 (T Lymphoblast, ATCC CRL-1582™) cell line were fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (dashed line histogram) or Alexa Fluor™ 647 Rat Anti-TOX antibody (solid line histogram) at 0.125 μg/test. The fluorescence histogram showing TOX expression (or Ig Isotype control staining) was derived from gated events with the side and forward light-scatter characteristics of intact MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this TDS are not lot specific.
Multiparameter flow cytometric analysis of TOX expression in mouse thymocytes. C57BL/6 mouse thymocytes were fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). Thymocytes were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561837/553048/553049), BD Horizon™ BV421 Rat Anti-Mouse CD8a (Cat. No. 563898) antibodies and with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731) or Alexa Fluor™ 647 Rat Anti-TOX antibody (Cat. No. 568356) at 0.125 μg/test. Overlapping, colorized TOX expression histograms (Right Plot) were generated for the corresponding colorized gated thymocyte subsets defined by the correlated expression of CD8a versus CD4 (Left Plot) as indicated: CD4+CD8a+ (Blue line histogram), CD4loCD8alo (Green histogram) and CD4+CD8alo (Purple histogram) with Ig Isotype control staining shown for CD4+CD8+ thymocytes (Black histogram). Flow cytometry and data analysis were performed on gated events with the side and forward light-scatter characteristics of intact thymocytes using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet (TDS) are not lot specific.
Flow cytometric analysis of TOX expression in human MOLT-4 cells. Cells from the human MOLT-4 (T Lymphoblast, ATCC CRL-1582™) cell line were fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). The cells were then stained with Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (dashed line histogram) or Alexa Fluor™ 647 Rat Anti-TOX antibody (solid line histogram) at 0.125 μg/test. The fluorescence histogram showing TOX expression (or Ig Isotype control staining) was derived from gated events with the side and forward light-scatter characteristics of intact MOLT-4 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this TDS are not lot specific.
Multiparameter flow cytometric analysis of TOX expression in mouse thymocytes. C57BL/6 mouse thymocytes were fixed and permeabilized using the Transcription Factor Buffer Set (Cat. No. 562574/562725). Thymocytes were stained with PE Rat Anti-Mouse CD4 (Cat. No. 561837/553048/553049), BD Horizon™ BV421 Rat Anti-Mouse CD8a (Cat. No. 563898) antibodies and with either Alexa Fluor™ 647 Rat IgG1, κ Isotype Control (Cat. No. 557731) or Alexa Fluor™ 647 Rat Anti-TOX antibody (Cat. No. 568356) at 0.125 μg/test. Overlapping, colorized TOX expression histograms (Right Plot) were generated for the corresponding colorized gated thymocyte subsets defined by the correlated expression of CD8a versus CD4 (Left Plot) as indicated: CD4+CD8a+ (Blue line histogram), CD4loCD8alo (Green histogram) and CD4+CD8alo (Purple histogram) with Ig Isotype control staining shown for CD4+CD8+ thymocytes (Black histogram). Flow cytometry and data analysis were performed on gated events with the side and forward light-scatter characteristics of intact thymocytes using a BD LSRFortessa™ X20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet (TDS) are not lot specific.
Product Details
BD Pharmingen™
TOX; TOX 1; TOX1
Human (QC Testing), Mouse (Tested in Development)
Rat IgG1, κ
Recombinant Human TOX Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing ≤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 unreacted dye was 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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Alexa Fluor™ is a trademark of Life Technologies Corporation.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
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
Alexa Fluor® 647 Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
557731
PE Rat Anti-Mouse CD4 RUO
Size
0.1 mg
Cat No.
553048
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BV421 Rat Anti-Mouse CD8a RUO
Size
50 µg
Cat No.
563898
568356 Rev. 2
Antibody Details
NAN448B
The NAN44B monoclonal antibody specifically binds to the thymocyte selection-associated protein high mobility group box protein, which is also known as TOX, TOX 1 and thymus high mobility group box protein. TOX belongs to the superfamily of transcription factors that contain the highly conserved high mobility group box (HMG-box) region, including a small subfamily of proteins (TOX2, TOX3 and TOX4) that share almost identical (HMG)-box sequences. TOX acts as a key regulator in the development of the adaptive immune system and is expressed by CD4+ helper T cells, CD8+cytotoxic T cells, regulatory T cells, NK cells, CD1D-dependent (NKT) cells and lymphoid tissue-inducer (LTi) cells. This DNA-binding factor regulates transcription by modifying chromatin structure and can reportedly serve as a critical regulator of T cell exhaustion and a potential immunotherapy target.
568356 Rev. 2
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
568356 Rev.2
Citations & References
View product citations for antibody "568356" on CiteAb
Development References (7)
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Aliahmad P, de la Torre B, Kaye J. Shared dependence on the DNA-binding factor TOX for the development of lymphoid tissue-inducer cell and NK cell lineages.. Nat Immunol. 2010; 11(10):945-52. (Biology). View Reference
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Jindal S, Pennock ND, Sun D, et al. Postpartum breast cancer has a distinct molecular profile that predicts poor outcomes.. Nat Commun. 2021; 12(1):6341. (Clone-specific). View Reference
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Liang C, Huang S, Zhao Y, Chen S, Li Y. TOX as a potential target for immunotherapy in lymphocytic malignancies.. Biomark Res. 2021; 9(1):20. (Biology). View Reference
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Maestre L, García-García JF, Jiménez S, et al. High-mobility group box (TOX) antibody a useful tool for the identification of B and T cell subpopulations.. PLoS One. 2020; 15(2):e0229743. (Clone-specific: Flow cytometry). View Reference
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Scott AC, Dündar F, Zumbo P, et al. TOX is a critical regulator of tumour-specific T cell differentiation.. Nature. 2019; 571(7764):270-274. (Biology). View Reference
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Veldman J, Rodrigues Plaça J, Chong L, et al. CD4+ T cells in classical Hodgkin lymphoma express exhaustion associated transcription factors TOX and TOX2: Characterizing CD4+ T cells in Hodgkin lymphoma.. Oncoimmunology. 2022; 11(1):2033433. (Clone-specific). View Reference
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Wilkinson B, Chen JY, Han P, Rufner KM, Goularte OD, Kaye J. TOX: an HMG box protein implicated in the regulation of thymocyte selection.. Nat Immunol. 2002; 3(3):272-80. (Biology). View Reference
568356 Rev. 2
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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.