Product Pathways - Growth Factors/Cytokines
Mouse Interferon-γ (mIFN-γ) #5222
PhosphoSitePlus® protein, site, and accession data: IFNG
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Source
Recombinant mouse IFN-γ (mIFN-γ) His23-Cys155 (Accession # NP_032363) was produced in E. coli at Cell Signaling Technology.
Molecular Characterization
Recombinant mIFN-γ has a Met on the amino terminus and has a calculated MW of 15,652. DTT-reduced and non-reduced protein migrate as 14 kDa polypeptides. The expected amino-terminus MHGTV of recombinant mIFN-γ was verified by amino acid sequencing.
Purity
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIFN-γ. All lots are greater than 98% pure.
Bioactivity
The bioactivity of mIFN-γ was determined in a virus protection assay. The ED50 of each lot is between 30-150 pg/ml.
Coomassie Gel
The purity of recombinant mIFN-γ was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIFN-γ and staining overnight with Coomassie Blue.
Bioactivity
The bioactivity of recombinant mIFN-γ was determined in a virus protection assay. L-929 cells were pretreated with increasing concentrations of mIFN-γ for 24 hours. Cells were then innoculated with encephalomyocarditis virus (EMCV) and incubated for an additional 48 hours. Surviving cells were then fixed and stained with crystal violet and the OD595 was determined.
Western Blotting
Western blot analysis of extracts from L-929 cells, untreated or treated with mIFN-γ for 20 minutes, using Phospho-Stat1 (Tyr701) Antibody #9171 (upper) and Stat1 Antibody #9172 (lower).
Endotoxin
Less than 0.01 ng endotoxin/1 μg mIFN-γ.
Formulation
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mIFN-γ. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Background
IFN-γ plays key roles in both the innate and adaptive immune response. IFN-γ activates the cytotoxic activity of innate immune cells such as macrophages and NK cells (1,2). IFN-γ production by NK cells and antigen-presenting cells (APCs) promotes cell-mediated adaptive immunity by inducing IFN-γ production by T lymphocytes, increased class I and class II MHC expression, and enhancing peptide antigen presentation (1). The anti-viral activity of IFN-γ is due to its induction of PKR and other regulatory proteins. Binding of IFN-γ to the IFNGR1/IFNGR2 complex promotes dimerization of the receptor complexes. Binding induces a conformational change in receptor intracellular domains and signaling involves Jak1, Jak2, and Stat1 (3). The critical role of IFN-γ in amplification of immune surveillance and function is supported by increased susceptibility to pathogen infection by IFN-γ or IFNGR knockout mice, and in humans with inactivating mutations in IFNGR1 or IFNGR2. IFN-γ also appears to have a role in atherosclerosis (4).
- Schroder, K. et al. (2004) J Leukoc Biol 75, 163-89.
- Martinez, F.O. et al. (2009) Annu Rev Immunol 27, 451-83.
- Kotenko, S.V. et al. (1995) J Biol Chem 270, 20915-21.
- McLaren, J.E. and Ramji, D.P. (2009) Cytokine Growth Factor Rev 20, 125-35.
Application References
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Companion Products
- 9171 Phospho-Stat1 (Tyr701) Antibody
- 9172 Stat1 Antibody
- 9872 Phosphate Buffered Saline (PBS-1X) pH7.2 (Sterile)
For Research Use Only. Not For Use In Diagnostic Procedures.
