Purified Mouse Anti-Human IFN-γ
Clone B27 (RUO)
- Brand BD Pharmingen™
- Alternative Name IFNG; Interferon-gamma; Interferon-γ; Type II interferon; MAF
- Concentration 0.5 mg/ml
- Isotype Mouse IgG1, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus, Baboon (Tested in Development)
ELISA (Routinely Tested)
Neutralization, Blocking (Tested During Development)
- Immunogen Human IFN-γ Recombinant Protein
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The B27 monoclonal antibody specifically binds to human interferon-γ (IFN-γ), a 14-18 kDa glycoprotein containing 143 amino acid residues. IFN-γ is a potent multifunctional cytokine 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 anti-tumor responses of macrophages, NK cells, and neutrophils. In addition, IFN-γ influences the regulation of 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 direct effects on B cells and T cells themselves. B27 is a neutralizing antibody. The use of B27 antibody for epitope mapping of human IFN-γ has been described. The B27 antibody has been reported not to bind to denatured IFN-γ.
Suggested Companion Products
Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
Blocking Control for Intracellular Staining: The purified B27 antibody can be used as a blocking control to demonstrate specificity of IFN-γ
staining by PE-B27 antibody or FITC-B27 antibody (Cat. No. 554701/554700). To perform this control, the fixed/permeabilized cells (~1
million) can be incubated with 1-10 µg of unlabeled B27 antibody (Cat. No. 554669) for 20 minutes at 4°C, prior to staining with PE-B27
antibody or FITC-B27 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.
Neutralization: The NA/LE™ B27 antibody is useful for neutralization of human IFN-γ bioactivity. This purified B27 antibody (Cat. No. 554698) is supplied in sodium azide free, sterile-filtered (0.22 µm pore) PBS, pH 7.2. Endotoxin level as determined by LAL assay is less than 0.01 ng/µg protein.
IP/WB: The B27 antibody has been reported to be useful for immunoprecipitation studies. The B27 antibody has been reported not to bind to denatured IFN-γ.