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      RY775 Rat Anti-Mouse IFN-γ
      RY775 Rat Anti-Mouse IFN-γ
      Two-color flow cytometric analysis of IFN-γ expression in stimulated Mouse splenic leukocytes.  BALB/c Mouse splenocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139; 50 ng/ml) and Ionomycin (Sigma I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either BD Horizon™ RY775 Rat IgG1, κ Isotype Control (Cat. No. 571678; Left Plot) or BD Horizon™ RY775 Rat Anti-Mouse IFN-γ antibody (Cat. No. 571701/571773; Right Plot) at 0.25 µg/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-γ (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      RY775 Rat Anti-Mouse IFN-γ
      Two-color flow cytometric analysis of IFN-γ expression in stimulated Mouse splenic leukocytes.  BALB/c Mouse splenocytes were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (PMA; Sigma P-8139; 50 ng/ml) and Ionomycin (Sigma I-0634; 1 μg/ml) in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) [Cat. No. 554724]. The cells were harvested and washed with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656] and fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either BD Horizon™ RY775 Rat IgG1, κ Isotype Control (Cat. No. 571678; Left Plot) or BD Horizon™ RY775 Rat Anti-Mouse IFN-γ antibody (Cat. No. 571701/571773; Right Plot) at 0.25 µg/test using BD Biosciences Intracellular Cytokine Staining protocol. Bivariate pseudocolor density plots showing the correlated expression of IFN-γ (or Ig Isotype control staining) versus CD4 were derived from gated events with the forward and side light-scatter characteristics of intact stimulated leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      商品详情
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      BD Horizon™
      Ifg; Ifng; IFN-γ; IFN-g; IFN-gamma; Interferon gamma; Type II Interferon
      Mouse (QC Testing)
      Rat IgG1, κ
      Mouse IFN-γ Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      15978
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      准备和存储

      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 unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      推荐的实验流程

      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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      商品通知

      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. 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.
      8. 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.
      9. 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.
      10. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      11. For U.S. patents that may apply, see bd.com/patents.
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      571773 Rev. 1
      抗体详情
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      XMG1.2

      The XMG1.2 monoclonal antibody specifically binds to mouse interferon-γ (IFN-γ) protein. IFN-γ is a pleiotropic cytokine, of approximately 15-17 kDa, involved in the regulation of inflammatory and immune responses. It plays an important role in activation, growth, and differentiation of T and B lymphocytes, macrophages, NK cells and other non-hematopoietic cell types. IFN-γ production is associated with the Th1 cell differentiation. The purified form of this antibody has been reported to be a neutralizing antibody.

      571773 Rev. 1
      格式详情
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      RY775
      The BD Horizon RealYellow™ 775 (RY775) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 775-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY775 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY775 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).
      altImg
      RY775
      Yellow-Green 561 nm
      557 nm
      775 nm
      571773 Rev.1
      报价单和参考
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      View product citations for antibody "571773" on CiteAb

      研发参考 (7)

      1. Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA, Neutralization). 查看参考
      2. Cherwinski HM, Schumacher JH, Brown KD, Mosmann TR. Two types of mouse helper T cell clone. III. Further differences in lymphokine synthesis between Th1 and Th2 clones revealed by RNA hybridization, functionally monospecific bioassays, and monoclonal antibodies. J Exp Med. 1987; 166(5):1229-1244. (Clone-specific: ELISA). 查看参考
      3. Klinman D and Nutman T. ELISPOT assay to detect cytokine-secreting murine and human cells. In: Coligan J, Kruisbeek A, Margulies D, Shevach E, Strober W, ed. Current Protocols in Immunology. 1994:6-19.
      4. Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology). 查看参考
      5. Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific: Intracellular Staining/Flow Cytometry). 查看参考
      6. Suzuki Y, Yang Q, Conley FK, Abrams JS, Remington JS. Antibody against interleukin-6 reduces inflammation and numbers of cysts in brains of mice with toxoplasmic encephalitis. Infect Immun. 1994; 62(7):2773-2778. (Clone-specific: In vivo exacerbation, Neutralization). 查看参考
      7. Yang X, HayGlass KT. A simple, sensitive, dual mAb based ELISA for murine gamma interferon determination: comparison with two common bioassays. J Immunoassay. 1993; 14(3):129-148. (Clone-specific: ELISA). 查看参考
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      571773 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.