Description
The GIR-94 antibody specifically recognizes the extracellular region of the alpha chain subunit (80-95 kDa glycoprotein) of the human interferon-γ receptor (IFN-γRα; aka, CD119). The functionally active-form of the human IFN-γ receptor consists of two (or more) subunits, with IFN-γRα responsible for IFN-γ binding and both the IFN-γR α and β chains required for the transduction of biologic responses. The IFN-γ receptor α chain (CD119) is expressed on the surface of most human cells (except mature erythrocytes) including monocytes, macrophages, T cells, B cells, NK cells, neutrophils, fibroblasts, epithelial cells, and endothelium. The ability of this antibody to bind to IFN-γ receptors of species other than human has not been determined. The immunogen used to generate this hybridoma was human IFN-γRα purified from human placenta. The GIR-94 is a non-neutralizing antibody.
Immunofluorescent staining and flow cytometric analysis: Purified Mouse Anti-Human CD119 (Cat. No. 558935) can be used for the immunofluorescent staining (≤ 1 µg/ 10[6] cells) and flow cytometric analysis of the levels of membrane IFN-γRα expressed by human cell lines or human lymphoid cells. It is recommended that the purified format of this antibody be used in conjunction with Biotin Goat Anti-Mouse Ig (Multiple Adsorption)(Cat No. 553999) and PE Streptavidin (Cat No. 554061) in a three-layer staining procedure to amplify immunofluorescent signals. An appropriate purified immunoglobulin isotype control is Purified Mouse IgG2b κ Isotype Control (Cat No. 555740). Since the GIR-94 is a non-neutralizing antibody, it can be used for immunofluorescent staining and flow cytometric analysis in systems where the IFN-γ ligand is present. Based on our testing results, the presence of recombinant human IFN-γ protein at levels below 1 µg/ml is insufficient to inhibit the binding of GIR-94 (at 0.06 µg mAb/1 million cells).
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
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. 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.
5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
