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FITC Rat Anti-Mouse IFN-γ XMG1.2 RUO

FITC Rat Anti-Mouse IFN-γ 

Clone  XMG1.2   (RUO)

  • Brand BD Pharmingen™
  • Alternative Name Ifg; Ifng; IFN-γ; IFN-g; IFN-gamma; Interferon gamma; Type II Interferon
  • Concentration 0.5 mg/ml
  • Isotype Rat IgG1, κ
  • Reactivity Mouse (QC Testing) 
  • Application Intracellular staining (flow cytometry) (Routinely Tested) 
  • Immunogen Mouse IFN-γ Recombinant Protein
  • Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
  • Regulatory Status RUO
Product Details Recommended Assay References

Description

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.

Format
  • Format FITC
  • Excitation Source Blue 488 nm
  • Excitation Max 494 nm
  • Emission Max 520 nm

FITC, fluorescein isothiocyanate, is a fluorochrome with a molecular weight of 389 Da. FITC is sensitive to pH changes and photobleaching. Due to nearly identical excitation and emission properties but different spillover characteristics, FITC and Alexa Fluor® 488 cannot be used simultaneously. FITC is relatively dim and should be reserved for highly expressed markers whenever possible.

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FITC Rat IgG1, κ Isotype Control RUO
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Purified NA/LE Hamster Anti-Mouse CD3e 145-2C11 RUO
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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 with FITC under optimum conditions, and unreacted FITC was removed.

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. 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.
  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.


Immunofluorescent Staining and Flow Cytometric Analysis: The FITC-conjugated XMG1.2 antibody can be used for multicolor immunofluorescent staining and flow cytometric analyses to identify and enumerate IFN-γ producing cells within mixed cell populations. For optimal immunofluorescent staining with flow cytometric analysis, this anti-cytokine antibody should be titrated (≤0.5 µg mAb/million cells).

A useful control for deomonstrating specificity of staining is either of the following: (1) pre-block the conjugated XMG1.2 antibody with a molar excess of ligand (e.g., recombinant mouse IFN-γ; Cat. No. 554587) prior to staining, or (2) pre-block the fixed/permeabilized cells with un-conjugated XMG1.2 antibody (Cat. No. 554409) prior to staining. The staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcelfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse cells is FITC-R3-34 (Cat. No. 554684); use at comparable concentrations to antibody of interest.


  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. 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. View reference

  3. Ferrick DA, Schrenzel MD, Mulvania T, Hsieh B, Ferlin WG, Lepper H. Differential production of interferon-gamma and interleukin-4 in response to Th1- and Th2-stimulating pathogens by gamma delta T cells in vivo. Nature. 1995; 373(6511):255-257. View reference

  4. Hsieh B, Schrenzel MD, Mulvania T, Lepper HD, DiMolfetto-Landon L, Ferrick DA. In vivo cytokine production in murine listeriosis. Evidence for immunoregulation by gamma delta+ T cells. J Immunol. 1996; 156(1):232-237. View reference

  5. 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. View reference

  6. 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. View reference

  7. Vikingsson A, Pederson K, Muller D. Enumeration of IFN-gamma producing lymphocytes by flow cytometry and correlation with quantitative measurement of IFN-gamma. J Immunol Methods. 1994; 173(2):219-228. View reference

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