Recombinant Human IFN-gamma GMP Protein, CF

Animal-Free.
  
  • Purity
    >97%, by SDS-PAGE with silver staining, under reducing conditions.
  • Endotoxin Level
    <0.01 EU per 1 μg of the protein by the LAL method.
  • 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 0.15-0.75 ng/mL.
    The specific activity of recombinant human IFN-gamma is approximately 2 x 104 IU/μg, which is calibrated against human IFN-gamma Standard (NIBSC code: 87/586).
  • Source
    E. coli-derived Gln24-Gln166, with an N-terminal Met Produced using non-animal reagents in an animal-free laboratory.
    Manufactured and tested under cGMP guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis
    Met-Gln24-Asp-Pro-Tyr-Val-Lys-Glu-Ala-Glu
  • Predicted Molecular Mass
    16.9 kDa
  • SDS-PAGE
    17 kDa, reducing conditions
285-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Succinate, Mannitol and Tween® 80.
Reconstitution Reconstitute at 200 μg/mL in sterile, deionized water.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images

GMP-grade Recombinant Human IFN-gamma (Catalog # 285-GMP) demonstrates anti-viral activity in HeLa human cervical epithelial carcinoma cells infected with encephalomyocarditis (EMC) virus. The ED50 for this effect is 0.15-0.75 ng/mL.

1 μg/lane of GMP-grade Recombinant Human IFN-gamma (Catalog # 285-GMP) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 17 kDa.

Mass Spectrometry

MALDI-TOF analysis of GMP-grade Recombinant Human IFN-gamma (Catalog #
285-GMP). The major peak corresponds to the calculated molecular mass, 16881 Da. The minor peak at 17099 Da is a matrix-associated artifact of the MALDI-TOF.

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). It shares 90% amino acid (aa) sequence identity with rhesus IFN-gamma, 59%‑64% with bovine, canine, equine, feline, and porcine IFN‑ gamma, and 37%‑43% with cotton rat, mouse, and rat IFN-gamma. 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 (4, 5). IFN-gamma is produced by a variety of immune cells under inflammatory conditions, notably by T cells and NK cells (6). 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 (6, 7). In addition, IFN-gamma functions as an anti-inflammatory mediator by promoting the development of regulatory T cells and inhibiting Th17 cell differentiation (8, 9). The pleiotropic effects of IFN-gamma contribute to the development of multiple aspects of atherosclerosis (7).
  • References:
    1. Billiau, A. and P. Matthys (2009) Cytokine Growth Factor Rev. 20:97.
    2. Pestka, S. et al. (2004) Immunol. Rev. 202:8.
    3. Gray, P.W. and D.V. Goeddel (1982) Nature 298:859.
    4. Marsters, S.A. et al. (1995) Proc. Natl. Acad. Sci. 92:5401.
    5. Krause, C.D. et al. (2000) J. Biol. Chem. 275:22995.
    6. Schroder, K. et al. (2004) J. Leukoc. Biol. 75:163.
    7. McLaren, J.E. and D.P. Ramji (2009) Cytokine Growth Factor Rev. 20:125.
    8. Muhl, H. and J. Pfeilschifter (2003) Int. Immunopharmacol. 3:1247.
    9. 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; IFNG; IFNgamma; IFN-gamma; Immune interferon; interferon gamma; interferon, gamma

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