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      Alexa Fluor® 700 Rat Anti-Mouse IFN-γ
      Alexa Fluor® 700 Rat Anti-Mouse IFN-γ
      IFN-γ expression in stimulated BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA (5 ng/ml, Sigma Cat. No. P-8139) and Ionomycin (500 ng, Sigma Cat. No. I-0634) in the presence of Brefeldin-A (GolgiPlug, Cat. No. 555029). Cells were harvested, fixed, permeabilized and stained with either rat anti-mouse IFG-γ antibody (Alexa Fluor® 700 XMG1.2, Cat. No. 557998), or immunoglobulin isotype control (Alexa Fluor® 700 R3-34, Cat. No. 558001) by using the BD Biosciences Pharmingen staining protocol.
      Alexa Fluor® 700 Rat Anti-Mouse IFN-γ
      IFN-γ expression in stimulated BALB/c spleen cells. Splenocytes from BALB/c mice were stimulated for 4 hours with PMA (5 ng/ml, Sigma Cat. No. P-8139) and Ionomycin (500 ng, Sigma Cat. No. I-0634) in the presence of Brefeldin-A (GolgiPlug, Cat. No. 555029). Cells were harvested, fixed, permeabilized and stained with either rat anti-mouse IFG-γ antibody (Alexa Fluor® 700 XMG1.2, Cat. No. 557998), or immunoglobulin isotype control (Alexa Fluor® 700 R3-34, Cat. No. 558001) by using the BD Biosciences Pharmingen staining protocol.
      Product Details
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      BD Pharmingen™
      Mouse (QC Testing)
      Rat IgG1, κ
      Mouse IFN-γ Recombinant Protein
      Intracellular staining (flow cytometry) (Routinely Tested)
      15-17 kDa
      0.2 mg/ml
      15978
      AB_396979
      Aqueous buffered solution containing protein stabilizer and ≤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 Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      Recommended Assay Procedure:

      Immunofluorescent Staining and Flow Cytometric Analysis: XMG1.2 is useful for immunofluorescent staining and flow cytometric analysis on fixed and permeabilized cells to identify and enumerate IFN-γ producing cells within mixed cell populations. An appropriate rat IgG1 isotype control is BD Cat. No. 558001. For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      3. Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. 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.
      6. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
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      557998 Rev. 2
      Antibody Details
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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.

      This antibody is routinely tested by flow cytometric analysis. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.

      557998 Rev. 2
      Format Details
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      Alexa Fluor™ 700
      Alexa Fluor™ 700 dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 697 nm and an emission maximum (Em Max) at 719-nm. Alexa Fluor™ 700 is designed to be excited by the Red (627–640-nm) laser and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 700
      Red 627-640 nm
      697 nm
      719 nm
      557998 Rev.2
      Citations & References
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      View product citations for antibody "557998" on CiteAb

      Development References (4)

      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). View Reference
      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). View Reference
      3. 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: Flow cytometry). View Reference
      4. 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). View Reference
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      557998 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.