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      BUV615 Rat Anti-Mouse CD357 (GITR)
      Product Details
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      BD OptiBuild™
      GITR; Gitr; glucocorticoid-induced TNFR-related protein; Tnfrsf18; AITR
      Mouse (Tested in Development)
      Rat IgG2b
      Mouse CD25+ CD4+ T Cell Line
      Flow cytometry (Qualified)
      0.2 mg/ml
      21936
      AB_2875527
      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 that minimize unconjugated dye and antibody.
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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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

      For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

      Note:  When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed.  For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).

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      Product Notices

      1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
      2. Researchers should determine the optimal concentration of this reagent for their individual applications.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      4. 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.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. CF™ is a trademark of Biotium, Inc.
      10. BD Horizon Brilliant Ultraviolet 615 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
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      751532 Rev. 2
      Antibody Details
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      DTA-1

      The DTA-1 monoclonal antibody specifically binds to GITR [Glucocorticoid-induced Tumor necrosis factor (TNF) receptor family-Related], a 66-70-kDa homodimer glycoprotein that is a member of the TNF receptor superfamily and is also known as TNFRSF18 and CD357. As its name implies, GITR expression was first detected in T lymphocytes that had been treated with dexamethasone, a glucocorticoid. In normal naive mice, GITR is expressed at moderate levels on CD25-positive/CD4-positive/CD8a-negative thymocytes and on CD25-positive/CD4-positive/CD45RB-low splenocytes. It is also expressed at low levels on splenic CD25-negative/CD4-positive/CD45RB-low T lymphocytes, B lymphocytes, macrophages, and dendritic cells. Activation of T and B lymphocytes upregulates GITR expression. GITR is a costimulatory receptor that plays an important role in Regulatory T (Treg)-cell functions, and a GITR Ligand has been detected on B lymphocytes, macrophages, and dendritic cells. mAb DTA-1 abrogates suppression by Treg cells without affecting their proliferative response, while it is co-stimulatory for T lymphocytes that are not Treg cells.

      The antibody was conjugated to BD Horizon BUV615 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome with an Ex Max near 350 nm and an Em Max near 615 nm. BD Horizon Brilliant BUV615 can be excited by the ultraviolet laser (355 nm) and detected with a 610/20 filter and a 595 nm LP.  Due to the excitation of the acceptor dye by the blue/yellow-green laser line, there may be significant spillover into channels detecting PE-CF594 like emissions (eg, 610/20-nm filter).

      751532 Rev. 2
      Format Details
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      BUV615
      The BD Horizon Brilliant™ Ultraviolet 615 (BUV615) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. BUV615, driven by BD innovation, is designed to be excited by the ultraviolet laser (355 nm) and detected using an optical filter centered near 615-nm (e.g, 610/20 bandpass filter). The acceptor dye can be excited by the Blue (488-nm) and yellow-green (561-nm) lasers resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      BUV615
      Ultraviolet 355 nm
      350 nm
      615 nm
      751532 Rev.2
      Citations & References
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      Development References (16)

      1. Beilharz MW, Sammels LM, Paun A, et al. Timed ablation of regulatory CD4-positive T cells can prevent murine AIDS progression. J Immunol. 2004; 172:4917-4925. (Biology). View Reference
      2. Beilharz MW, Sammels LM, Paun A, et al. Timed ablation of regulatory CD4-positive T cells can prevent murine AIDS progression. J Immunol. 2004; 172:4917-4925. (Clone-specific). View Reference
      3. Dittmer U, He H, Messer RJ, et al. Functional impairment of CD8-positive T cells by regulatory T cells during persistent retroviral infection. Immunity. 2004; 20:293-303. (Clone-specific). View Reference
      4. Dittmer U, He H, Messer RJ, et al. Functional impairment of CD8-positive T cells by regulatory T cells during persistent retroviral infection. Immunity. 2004; 20:293-303. (Clone-specific: In vivo exacerbation). View Reference
      5. Ji H, Liao G, Faubion WA, et al. The natural ligand for glucocorticoid-induced TNF receptor-related protein abrogates regulatory T cell suppression. J Immunol. 2004; 172:5823-5827. (Biology). View Reference
      6. Kohm AP, Williams JS, Miller SD. Ligation of the glucocorticoid-induced TNF receptor enhances autoreactive CD4-positive T cell activation and experimental autoimmune encephalomyelitis. J Immunol. 2004; 172:4686-4690. (Clone-specific). View Reference
      7. Kohm AP, Williams JS, Miller SD. Ligation of the glucocorticoid-induced TNF receptor enhances autoreactive CD4-positive T cell activation and experimental autoimmune encephalomyelitis. J Immunol. 2004; 172:4686-4690. (Clone-specific: Flow cytometry, In vivo exacerbation). View Reference
      8. Nocentini G, Giunchi L, Ronchetti S, et al. A new member of the tumor necrosis factor/nerve growth factor receptor family inhibits T cell receptor-induced apoptosis. Proc Natl Acad Sci U S A. 1997; 94:6216-6221. (Biology). View Reference
      9. Nocentini G, Giunchi L, Ronchetti S, et al. A new member of the tumor necrosis factor/nerve growth factor receptor family inhibits T cell receptor-induced apoptosis. Proc Natl Acad Sci U S A. 1997; 94:6216-6221. (Biology). View Reference
      10. Shimizu J, Moriizumi E. CD4-positive CD25-negative T cells in aged mice are hyporesponsive and exihibit suppressive activity. J Immunol. 2003; 170:1675-1682. (Clone-specific). View Reference
      11. Shimizu J, Moriizumi E. CD4-positive CD25-negative T cells in aged mice are hyporesponsive and exihibit suppressive activity. J Immunol. 2003; 170:1675-1682. (Clone-specific: Functional assay). View Reference
      12. Shimizu J, Yamazaki S, Takahashi T, Ishida Y, Sakaguchi S. Stimulation of CD25-positive CD4-positive regulatory T cells through GITR breaks immunological self-tolerance. Nat Immunol. 2002; 3(2):135-142. (Immunogen). View Reference
      13. Shimizu J, Yamazaki S, Takahashi T, Ishida Y, Sakaguchi S. Stimulation of CD25-positive CD4-positive regulatory T cells through GITR breaks immunological self-tolerance. Nat Immunol. 2002; 3(2):135-142. (Immunogen: Cell separation, Depletion, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition, In vivo exacerbation). View Reference
      14. Tone M, Tone Y, Adams E, et al. Mouse glucocorticoid-induced tumor necrosis factor receptor ligand is costimulatory for T cells. Proc Natl Acad Sci U S A. 2003; 100(25):15059-15064. (Biology). View Reference
      15. Uraushihara K, Kanai T, Ko K, et al. Regulation of murine inflammatory bowel disease by CD25-positive and CD25negative CD4-positive glucocorticoid-induced TNF receptor family-related gene-positive regulatory T cells. J Immunol. 2003; 171:708-716. (Clone-specific). View Reference
      16. Uraushihara K, Kanai T, Ko K, et al. Regulation of murine inflammatory bowel disease by CD25-positive and CD25negative CD4-positive glucocorticoid-induced TNF receptor family-related gene-positive regulatory T cells. J Immunol. 2003; 171:708-716. (Clone-specific: Cell separation, Flow cytometry, In vivo exacerbation). View Reference
      View All (16) View Less
      751532 Rev. 2

      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.

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