Purification notesAffinity isolated antigen specific antibody is obtained from goat anti-IFNGRI antiserum by immuno-specific purification which removes essentially all goat serum proteins, including immunoglobulins, which do not specifically bind to the peptide.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesELISA: Use at a concentration of 0.5 - 1 µg/ml. The detection limit for recombinant human IFNGRI is approximately 0.3 ng/well.
Neut: Use at a concentration of 0.5 - 2 µg/ml.
WB: Use at a concentration of 0.1 - 0.2 µg/ml. The detection limit for recombinant human IFNGRI is approximately 5 ng/lane under non-reducing and reducing conditions. Predicted molecular weight: 50 kDa.
To measure the ability of this antibody to block the cell surface recombinant human IFNGRI mediated bioactivity on HeLa cells, various concentrations of the antibody are added to confluent cultures of HeLa cells in a 96 well plate. The assay mixture in a total volume of 150 µl, containing antibody at concentrations from 0.01 to 50 µg/ml, is incubated for 1 hour at 37 °C. Recombinant human IFNG is added at 2 ng/ml and the mixture is incubated at 37 °C for 20 to 24 hours in a humidified CO2 incubator. At the end of this incubation period, the medium is aspirated from the wells and a titrated amount of Encephalomyocarditis virus (EMCV) in prewarmed culture medium is added to each test well. After an additional 20 to 24 hour incubation period, the cells are fixed, stained, and scored for cytopathic effect by measurement of optical densities in a microplate reader at 540 nm.
The Neutralization Dose50 (ND50) for anti-human IFNG receptor I is approximately 0.5 to 2.0 µg/ml in the presence of 2 ng/ml of recombinant human IFNG, using a confluent culture of HeLa cells. The ND50 is the concentration of antibody required to yield one-half maximal inhibition of the cytokine activity on a responsive cell line, when the cytokine is present at a concentration just high enough to elicit a maximum response. The exact concentration of antibody required to neutralize recombinant human IFNGRI activity is dependent on the cytokine concentration, cell type, growth conditions, and the type of activity studied.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionReceptor for interferon gamma. Two receptors bind one interferon gamma dimer.
Involvement in diseaseDefects in IFNGR1 are a cause of mendelian susceptibility to mycobacterial disease (MSMD) [MIM:209950]; also known as familial disseminated atypical mycobacterial infection. This rare condition confers predisposition to illness caused by moderately virulent mycobacterial species, such as Bacillus Calmette-Guerin (BCG) vaccine and environmental non-tuberculous mycobacteria, and by the more virulent Mycobacterium tuberculosis. Other microorganisms rarely cause severe clinical disease in individuals with susceptibility to mycobacterial infections, with the exception of Salmonella which infects less than 50% of these individuals. The pathogenic mechanism underlying MSMD is the impairment of interferon-gamma mediated immunity whose severity determines the clinical outcome. Some patients die of overwhelming mycobacterial disease with lepromatous-like lesions in early childhood, whereas others develop, later in life, disseminated but curable infections with tuberculoid granulomas. MSMD is a genetically heterogeneous disease with autosomal recessive, autosomal dominant or X-linked inheritance.
Sequence similaritiesBelongs to the type II cytokine receptor family. Contains 2 fibronectin type-III domains. Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
Post-translational modificationsPhosphorylated at Ser/Thr residues.