BD Horizon™ RB780 Rat Anti-TOX
Clone NAN448B (RUO)
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 then stained with BD OptiBuild™ BV421 Rat Anti-Mouse CD4 (Cat. No. 740007), PE Rat Anti-Mouse CD8a (Cat. No. 553033) antibodies and with either BD Horizon™ RB780 Rat IgG1, κ Isotype Control (Cat. No. 570465) or BD Horizon™ RB780 Rat Anti-TOX antibody (Cat. No. 570192/570193) at 0.25 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™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of TOX expression in Human MOLT-4 cells. Cells from the Human MOLT-4 (T lymphoblastic leukemia, 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 BD Horizon™ RB780 Rat IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ RB780 Rat Anti-TOX antibody (solid line histogram) at 0.25 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™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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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 then stained with BD OptiBuild™ BV421 Rat Anti-Mouse CD4 (Cat. No. 740007), PE Rat Anti-Mouse CD8a (Cat. No. 553033) antibodies and with either BD Horizon™ RB780 Rat IgG1, κ Isotype Control (Cat. No. 570465) or BD Horizon™ RB780 Rat Anti-TOX antibody (Cat. No. 570192/570193) at 0.25 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™ X-20 Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
Flow cytometric analysis of TOX expression in Human MOLT-4 cells. Cells from the Human MOLT-4 (T lymphoblastic leukemia, 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 BD Horizon™ RB780 Rat IgG1, κ Isotype Control (dashed line histogram) or BD Horizon™ RB780 Rat Anti-TOX antibody (solid line histogram) at 0.25 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™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Horizon™
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
9760, 252838
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- 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.
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- 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).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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
RB780 Rat IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
570465
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BV421 Rat Anti-Mouse CD4 RUO
Size
50 µg
Cat No.
740007
PE Rat Anti-Mouse CD8a RUO
Size
0.2 mg
Cat No.
553033
570193 Rev. 1
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.
570193 Rev. 1
Format Details
RB780
The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
RB780
Blue 488 nm
498 nm
781 nm
570193 Rev.1
Citations & References
View product citations for antibody "570193" on CiteAb
Development References (5)
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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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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. (Immunogen: Immunofluorescence, Immunohistochemistry, Western blot). 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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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
570193 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.