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Purified Mouse Anti-Human IFN-γ
Purified Mouse Anti-Human IFN-γ
Flow cytometric analysis of IFN-γ expression in stimulated human peripheral blood mononuclear cells. HiCK-1 Human Cytokine Positive Control Cells (Cat. No. 555061) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either Purified Mouse IgG1, κ Isotype Control (Cat No. 556648, Left Panel) or with the Purified Mouse Anti-Human IFN-γ antibody (Cat No. 554549, Right Panel), followed by FITC Goat Anti-Mouse Ig (Cat. No. 554001). Flow cytometric dot plots showing the expression of IFN-γ (or Ig Isotype Control staining) versus side scatter were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ System.
Flow cytometric analysis of IFN-γ expression in stimulated human peripheral blood mononuclear cells. HiCK-1 Human Cytokine Positive Control Cells (Cat. No. 555061) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with either Purified Mouse IgG1, κ Isotype Control (Cat No. 556648, Left Panel) or with the Purified Mouse Anti-Human IFN-γ antibody (Cat No. 554549, Right Panel), followed by FITC Goat Anti-Mouse Ig (Cat. No. 554001). Flow cytometric dot plots showing the expression of IFN-γ (or Ig Isotype Control staining) versus side scatter were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD FACSCanto™ System.
Product Details
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BD Pharmingen™
IFNG; Interferon-gamma; IFG; IFI; Type II interferon
Human (QC Testing), Rhesus, Cynomolgus, Baboon (Tested in Development)
Mouse BALB/c IgG1, κ
Human IFN-γ from supernatants of S. aureus-stimulated PBMC
Intracellular block/flow cytometry (Routinely Tested), ELISA Detection, Western blot (Reported)
0.5 mg/ml
3458
AB_398569
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Recommended Assay Procedures

Blocking Control for Intracellular Staining: Purified Mouse Anti-Human IFN-γ antibody (Cat. No. 554549) can be used as a blocking control to demonstrate specificity of IFN-γ staining by directly conjugated clone 4S.B3. To perform this control, the fixed/permeabilized cells (~1 million) can be incubated with 1-10 µg of Purified Mouse Anti-Human IFN-γ for 20 minutes at 4°C, prior to staining with the directly conjugated antibody (e.g., 0.1 -0.5 µg mAb/1 million cells). The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. For specific methodology, please visit our web site, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "Intracellular Staining".

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  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. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  5. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
554549 Rev. 2
Antibody Details
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4S.B3

The 4S.B3 monoclonal antibody specifically binds to interferon-γ (IFN-γ). The immunogen used to generate this hybridoma was partially purified human IFN-γ obtained from supernatants of human PBMC stimulated with Staphylococcus aureus. Interferon-γ (IFN-γ) is a potent multifunctional cytokine that is produced by several activated cell types including NK, NKT, CD4+TCRαβ+, CD8+TCRαβ+, and TCRγδ+ T cells. IFN-γ exerts its biological effects through specific binding to the high-affinity IFN-γ Receptor Complex comprised of IFN-γRα (CD119) and IFN-γRβ subunits. In addition to its antiviral effects, IFN-γ upregulates a number of lymphoid cell functions including the antimicrobial and antitumor responses of macrophages, NK cells, and neutrophils. In addition, IFN-γ can exert strong regulatory influences on the proliferation, differentiation, and effector responses of B cell and T cell subsets. These influences can involve IFN-γ's capacity to boost MHC class I and II expression by antigen-presenting cells as well as to direct effects on B cells and T cells themselves. Human IFN-γ is a 14-18 kDa glycoprotein containing 143 amino acid residues.

Clone 4S.B3 also cross-reacts with a cytoplasmic component of peripheral blood CD3+ lymphocytes of baboon, and both rhesus and cynomolgus macaque monkeys following five-hour treatment with phorbol myristic acetate (PMA) and Ca++ ionophore (A23187) in the presence of monensin. The staining pattern of 4S.B3 in CD3+ cells is similar to that observed with peripheral blood T lymphocytes from normal human donors. This reagent is useful for intracellular immunofluorescent staining for flow cytometric analysis to identify and enumerate IFN-γ + cells within a mixed cell population.

554549 Rev. 2
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
554549 Rev.2
Citations & References
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Development References (3)

  1. Meager A, Parti S, Barwick S, Spragg J, O'Hagan K. Detection of hybridomas secreting monoclonal antibodies to human gamma interferon using a rapid screening technique and specificity of certain monoclonal antibodies to gamma interferon. J Interferon Res. 1984; 4(4):619-625. (Biology). View Reference
  2. Meager A. Characterization of interferons and immunoassays. In: Clemens MJ, Morris AG, Gearing AJH, ed. Lymphokines and Interferons. A Practical Approach. Oxford: IRL Press Ltd; 1987:105-127.
  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). View Reference
554549 Rev. 2

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