Biotin Mouse Anti-Human IFN-γ
Clone 4S.B3 (RUO)
- Brand BD Pharmingen™
- Alternative Name IFNG; Interferon-gamma; IFG; IFI; Type II interferon
- Concentration 0.5 mg/ml
- Isotype Mouse BALB/c IgG1, κ
- Reactivity Human (QC Testing) Rhesus, Cynomolgus, Baboon (Tested in Development)
ELISA Detection (Routinely Tested)
Immunofluorescence, Flow cytometry (Tested During Development)
Western blot (Reported)
- Immunogen Human IFN-γ from supernatants of S. aureus-stimulated PBMC
- Storage Buffer Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
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.
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Preparation and Storage
Store undiluted at 4°C.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
- 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.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
ELISA Detection: Biotin Mouse Anti-Human IFN-γ (Cat. No. 554550) is useful as a detection antibody in a sandwich ELISA for measuring human IFN-ã protein levels. Biotin Mouse Anti-Human IFN-ã can be paired with Purified Mouse Anti-Human IFN-γ (Cat. No. 551221) as the capture antibody, with Recombinant Human IFN-ã (Cat. No. 554616/554617) as the standard. Biotin Mouse Anti-Human IFN-γ should be titrated (0.5 -2.0 µg/ml) to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of human IFN-γ ranging from ~15 to 2,000 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "ELISA and ELISPOT."
Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines or chemokines tested (e.g., mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, G-CSF, GM-CSF, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23 , SCF, TNF, LT-α, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF).
Note 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring human IFN-γ in serum or plasma our Human IFN-g ELISA Set (Cat. No. 555142) or Human IFN-g ELISA Kit II (Cat. No. 550612) are specially formulated and recommended.
Immunofluorescent Staining and Flow Cytometric Analysis: The 4S.B3 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IFN-γ producing cells within mixed cell populations. FITC Mouse Anti-Human IFN-γ and PE Mouse Anti-Human IFN-γ (Cat. No. 554551/554552) are especially suitable for these studies. The use of a specificity control, such as one of the following, is suggested: 1) Recombinant Human IFN-γ (Cat. No. 554616), 2) unlabeled Purified Mouse Anti-Human IFN-γ (Cat. No. 554549), or 3) FITC Mouse IgG1 κ Isotype Control (Cat. No. 554679) and PE Mouse IgG1, κ Isotype Control (Cat. No. 554680).
Western Blot: The 4S.B3 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.