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      RB613 Mouse Anti-Human FoxP3
      RB613 Mouse Anti-Human FoxP3
      Multicolor flow cytometric analysis of FoxP3 expression in Human lymphocytes.  Human peripheral blood mononuclear cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424; Top Plots) and Alexa Fluor™ 700 Anti-Human CD25 (Cat. No. 561398; Bottom Plots) antibodies. The cells were washed, fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). The cells were then stained with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human FoxP3 antibody (Cat. No. 571236/571300; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of FoxP3 [or Ig Isotype control staining] versus either CD4 or CD25 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      RB613 Mouse Anti-Human FoxP3
      Multicolor flow cytometric analysis of FoxP3 expression in Human lymphocytes.  Human peripheral blood mononuclear cells were stained with BD Horizon™ BV421 Mouse Anti-Human CD4 (Cat. No. 562424; Top Plots) and Alexa Fluor™ 700 Anti-Human CD25 (Cat. No. 561398; Bottom Plots) antibodies. The cells were washed, fixed and permeabilized using the BD Pharmingen™ Transcription Factor Buffer Set (Cat. No. 562574). The cells were then stained with either BD Horizon™ RB613 Mouse IgG1, κ Isotype Control (Cat. No. 571106; Left Plots) or BD Horizon™ RB613 Mouse Anti-Human FoxP3 antibody (Cat. No. 571236/571300; Right Plots). Bivariate pseudocolor density plots showing the correlated expression of FoxP3 [or Ig Isotype control staining] versus either CD4 or CD25 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ Software.
      Product Details
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      BD Horizon™
      Forkhead box protein P3; Scurfin; AIID; IPEX; JM2; DIETER; PIDX; XPID
      Human (QC Testing)
      Mouse BALB/c IgG1, κ
      Human full-length FoxP3 Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      5 µl/test
      50943
      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.
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      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.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. 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.
      3. 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).
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. An isotype control should be used at the same concentration as the antibody of interest.
      6. CF™ is a trademark of Biotium, Inc.
      7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. 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.
      9. 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.
      10. 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.
      11. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
      12. For U.S. patents that may apply, see bd.com/patents.
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      571300 Rev. 1
      Antibody Details
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      236A/E7

      The 236A/E7 antibody reacts with the human FoxP3 (Forkhead box protein P3) transcription factor, a member of the forkhead or winged helix family of transcription factors. The expression of FoxP3, also known as Scurfin, IPEX and JM2, has been found to be associated with CD4+ CD25+ regulatory T cells and represents a specific marker for these cells. Flow cytometric analysis has shown that FoxP3 is expressed by the majority of CD4+ CD25+ (high) T cells in peripheral blood while less than half of CD4+ CD25+ (intermediate) cell population are FoxP3 positive. Approximately 5-10% of peripheral CD4+ cells are CD4+ CD25+ T regulatory cells. T regulatory cells are thought to play a critical role in the control of T cell mediated autoimmunity by suppressing the proliferation and cytokine production of other T cells. To support this hypothesis, it has been found that Foxp3 is mutated in scurfy (sf) mice. Cumulative evidence suggests that the 236A/E7 antibody recognizes epitopes from both isoforms of Exon 2 alternatively-spliced variants and is located in the region between Exon 2 and the Zn/LZ domains (aa 105-235).

      571300 Rev. 1
      Format Details
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      RB613
      The BD Horizon RealBlue™ 613 (RB613) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 492-nm and an emission maximum (Em Max) at 613-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB613 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with reduced excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB613 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-CF594. RB613 is on average brighter than PE-CF594 off the blue laser.
      altImg
      RB613
      Blue 488 nm
      492 nm
      613 nm
      571300 Rev.1
      Citations & References
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      View product citations for antibody "571300" on CiteAb

      Development References (8)

      1. Alvaro T, Lejeune M, Salvado MT, et al. Outcome in Hodgkin's lymphoma can be predicted from the presence of accompanying cytotoxic and regulatory T cells. Clin Cancer Res. 2005; 11(4):1467-1473. (Clone-specific: Immunohistochemistry). View Reference
      2. Brunkow ME, Jeffery EW, Hjerrild KA, et al. Disruption of a new forkhead/winged-helix protein, scurfin, results in the fatal lymphoproliferative disorder of the scurfy mouse. Nat Genet. 2001; 27(1):68-73. (Biology). View Reference
      3. Fox BC, Bignone PA, Brown PJ, Banham AH. Defense of the clone: antibody 259D effectively labels human FOXP3 in a variety of applications. Blood. 2008; 111(7):3897-3899. (Clone-specific: Western blot). View Reference
      4. Lennon G, Auffray C, Polymeropoulos M, Soares MB. Consortium: an integrated molecular analysis of genomes and their expression. Genomics. 1996; 33(1):151-152. (Biology). View Reference
      5. Roncador G, Brown PJ, Maestre L, et al. Analysis of FOXP3 protein expression in human CD4+CD25+ regulatory T cells at the single-cell level. Eur J Immunol. 2005; 35(6):1681-1691. (Immunogen: Flow cytometry, Fluorescence activated cell sorting, Immunohistochemistry). View Reference
      6. Roncador G, Garcia JF, Maestre L, et al. FOXP3, a selective marker for a subset of adult T-cell leukaemia/lymphoma. Leukemia. 2005; 19(12):2247-2253. (Immunogen: Immunohistochemistry). View Reference
      7. Wildin RS, Ramsdell F, Peake J, et al. X-linked neonatal diabetes mellitus, enteropathy and endocrinopathy syndrome is the human equivalent of mouse scurfy. Nat Genet. 2001; 27(1):18-20. (Biology). View Reference
      8. Wolf D, Wolf AM, Rumpold H, et al. The expression of the regulatory T cell-specific forkhead box transcription factor FoxP3 is associated with poor prognosis in ovarian cancer. Clin Cancer Res. 2005; 11(23):8326-8331. (Biology). View Reference
      View All (8) View Less
      571300 Rev. 1

       

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      For Research Use Only. Not for use in diagnostic or therapeutic procedures.