Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Interferon-γ (hIFN-γ) #8901

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Source

Recombinant human IFN-γ (hIFN-γ) Gln24-Gln166 (Accession # NM_000619) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Recombinant hIFN-γ has a Met on the amino terminus and has a calculated MW of 16,907. DTT-reduced and non-reduced protein migrate as 17 kDa polypeptides. The expected amino-terminus MQDPY of recombinant hIFN-γ was verified by amino acid sequencing.

Purity

>96% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIFN-γ. All lots are greater than 96% pure.

Bioactivity

The bioactivity of hIFN-γ was determined in a virus protection assay. The ED50 of each lot is between 0.30-1.2 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hIFN-γ was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIFN-γ and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The bioactivity of recombinant hIFN-γ was determined in a virus protection assay. HeLa cells were pretretated with increasing concentrations of hIFN-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 Blotting

Western blot analysis of extracts from HeLa cells, untreated or treated with hIFN-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 hIFN-γ.

Formulation

With carrier: A 0.22 μm filtered solution of 0.33 mg/ml hIFN-γ in PBS, pH 7.2 containing 20 μg BSA per 1 μg hIFN-γ. Carrier free: A 0.22 μm filtered solution of 0.33 mg/ml hIFN-γ in 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 the 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 to form (IFNGR1/IFNGR2)2 -IFN-γ dimer. 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).

  1. Schroder, K. et al. (2004) J Leukoc Biol 75, 163-89.
  2. Martinez, F.O. et al. (2009) Annu Rev Immunol 27, 451-83.
  3. Kotenko, S.V. et al. (1995) J Biol Chem 270, 20915-21.
  4. McLaren, J.E. and Ramji, D.P. (2009) Cytokine Growth Factor Rev 20, 125-35.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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