Source	Human embryonic kidney cell, HEK293-derived human IFN-gamma protein Met1-Gln166
Purity	>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin level	<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis	Gln24 inferred from enzymatic pyroglutamate treatment revealing Asp25.
Predicted Molecular Mass	16.8 kDa
SDS-PAGE	19-25 kDa, reducing conditions
Activity	Measured in anti-viral assays using HeLa human cervical epithelial carcinoma cells infected with encephalomyocarditis (EMC) virus. Meager, A. (1987) in Lymphokines and Interferons, a Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 129. The ED₅₀ for this effect is 0.06-0.36 ng/mL.

Recombinant Human IFN-gamma (HEK293-expressed) Protein, CF Scientific Data Examples

	Recombinant Human IFN-gamma (HEK293-expressed) Protein Bioactivity Recombinant Human IFN-gamma (Catalog # 10067‑IF) demonstrates anti-viral activity in HeLa human cervical epithelial carcinoma cells infected with encephalomyocarditis (EMC) virus. The ED₅₀ for this effect is 0.06‑0.36 ng/mL
	Recombinant Human IFN-gamma (HEK293-expressed) Protein SDS-PAGE 2 μg/lane of Recombinant Human IFN‑ gamma (HEK293-expressed) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 19-25 kDa.

Formulation, Preparation and Storage

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

10067-IF

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 250 μg/mL in PBS.
Shipping	The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, ≤ -20 °C under sterile conditions after reconstitution.

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: IFN-gamma

Interferon-gamma (IFN-gamma), also known as type II or immune interferon, exerts a wide range of immunoregulatory activities and is considered to be the prototype proinflammatory cytokine (1, 2). Mature human IFN-gamma exists as a non-covalently linked homodimer of 20-25 kDa variably glycosylated subunits (3). Glycosylation of IFN-gamma at sites Asn25 and Asn97 is critical for protease resistance (4, 5). The mature protein shares 36% and 35% amino acid (aa) sequence identity with mouse and rat IFN-gamma, respectively. IFN-gamma dimers bind to IFN-gamma RI (alpha subunits) which then interact with IFN-gamma RII (beta subunits) to form the functional receptor complex of two alpha and two beta subunits. Inclusion of IFN-gamma RII increases the binding affinity for ligand and the efficiency of signal transduction (6, 7). IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells (8). It plays a key role in host defense by promoting the development and activation of Th1 cells, chemoattraction and activation of monocytes and macrophages, up‑regulation of antigen presentation molecules, and immunoglobulin class switching in B cells. It also exhibits antiviral, antiproliferative, and apoptotic effects (8, 9). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (10, 11). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (9).

Item	Value
References	Billiau, A. and P. Matthys (2009) Cytokine Growth Factor Rev. 20:97. Pestka, S. et al. (2004) Immunol. Rev. 202:8. Gray, P.W. and D.V. Goeddel (1982) Nature 298:859. Sareneva, T. et al. (1994) J. Biochem. 303:831. Sareneva, T. et al. (1995) J. Biochem. 308:9. Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. 92:5401. Krause, C.D. et al. (2000) J. Biol. Chem. 275:22995. Schroder, K. et al. (2004) J. Leukoc. Biol. 75:163. McLaren, J.E. and D.P. Ramji (2009) Cytokine Growth Factor Rev. 20:125. Muhl, H. and J. Pfeilschifter (2003) Int. Immunopharmacol. 3:1247. Kelchtermans, H. et al. (2008) Trends Immunol. 29:479.
Long Name	Interferon gamma
Entrez Gene IDs	3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)
Alternate Names	IFG, IFI, IFN-gamma, IFNG, IFNgamma, Immune interferon, interferon gamma, interferon, gamma

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Recombinant Human IFN-gamma (HEK293-expressed) Protein, CF

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