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Recombinant Human IFN-gamma, Animal-Free Protein

Bio-Techne includes R&D Systems | Catalog # AFL285

Analyzed by SEC-MALS. Intended for preclinical researchers who may transition to GMP IFN-gamma for their clinical work
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AFL285
AFL285-100

Key Product Details

Source

E. coli

Accession #

Conjugate

Unconjugated

Applications

Bioactivity

Product Specifications

Source

E. coli-derived human IFN-gamma protein
Gln24-Gln166, with an N-terminal Met
Based on MALDI-TOF mass spectrometric analysis, truncation of four amino acid residues from the C-terminus may be observed in the recombinant protein preparation.
Produced using non-animal reagents in an animal-free laboratory.

Purity

>97%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.01 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

16.9 kDa

SDS-PAGE

17 and 16 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 0.15-0.75 ng/mL.
The specific activity of recombinant human IFN-gamma is >2.0 x 107 IU/mg, which is calibrated against the human IFN-gamma Standard (NIBSC code: 87/586).

Scientific Data Images for Recombinant Human IFN-gamma, Animal-Free Protein

Animal-Free™ Recombinant Human IFN‑ gamma Protein SEC-MALS.

Recombinant Human IFN-gamma (Catalog # AFL285) has a molecular weight (MW) of 34.1 kDa as analyzed by SEC-MALS, suggesting that this protein is a homodimer.  MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Formulation, Preparation and Storage

AFL285
Formulation Lyophilized from a 0.2 μm filtered solution in Sodium Succinate, Mannitol and Tween® 80.
Reconstitution Reconstitute at 0.2 mg/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.
  • 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 to -70 °C under sterile conditions after reconstitution.

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

Alternate Names

IFG, IFI, IFNG, IFNgamma

Entrez Gene IDs

3458 (Human); 15978 (Mouse); 25712 (Rat); 396991 (Porcine); 281237 (Bovine); 403801 (Canine); 493965 (Feline)

Gene Symbol

IFNG

UniProt

Additional IFN-gamma Products

Product Documents for Recombinant Human IFN-gamma, Animal-Free Protein

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Manufacturing Specifications

Animal-Free Manufacturing Conditions
Our dedicated controlled-access animal-free laboratories ensure that at no point in production are the products exposed to potential contamination by animal components or byproducts. Every stage of manufacturing is conducted in compliance with R&D Systems' stringent Standard Operating Procedures (SOPs). Production and purification procedures use equipment and media that are confirmed animal-free.  

 Production

  • All molecular biology procedures use animal-free media and dedicated labware.
  • Dedicated fermentors are utilized in committed animal-free areas.

Purification

  • Protein purification columns are animal-free.
  • Bulk proteins are filtered using animal-free filters.
  • Purified proteins are stored in animal-free containers in a dedicated cold storage room.

    Quality Assurance

    • Low Endotoxin Level.
    • No impairment of biological activity.
    • High quality product obtained under stringent conditions.
    • For ex vivo research or bioproduction, additional documentation can be provided.

    Please read our complete Animal-Free Statement

    Product Specific Notices for Recombinant Human IFN-gamma, Animal-Free Protein

    For research use or further manufacturing only

    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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