Recombinant Human IL-18/IL-1F4 GMP Protein, CF New

Catalog #: BT-018-GMP Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
BT-018-GMP-050
BT-018-GMP-01M

Save 15% on Select RUO Reagents. See Details.

Recombinant Human IL-18/IL-1F4 GMP Protein Bioactivity.

5 Images

Product Details

Reviews

Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human IL-18/IL-1F4 GMP Protein, CF Summary

IL-18 Manufactured in Bio-Techne's new GMP facility
Lot-to-lot consistency
Stringent guidelines for patient safety
Scalability necessary to support successful therapeutics
Learn more about manufacturing in our new GMP facility
Test it in your process! Request a sample of GMP IL-18

Product Specifications

Purity

>97%, by SDS-PAGE with quantitative densitometry by Coomassie® Blue Staining. The molecular weight by mass spectrometry is 18205 Da ± 5 Da

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

Measured by its ability to induce IFN-gamma secretion by KG‑1 human acute myelogenous leukemia cells in the presence of TNF-alpha. The ED₅₀ for this effect is 1.25-15.0 ng/mL.

The specific activity of GMP Recombinant Human IL-18/IL-1F4 is > 5.00 x 10⁶ units/mg, which is calibrated against the human IL-18 WHO Standard (NIBSC code: 03/200).

Source

E. coli-derived human IL-18/IL-1F4 protein
Tyr37-Asp193
Produced using non-animal reagents in an animal-free laboratory.

Manufactured and tested under cGMP guidlines.

Accession #

Q14116.1

N-terminal Sequence
Analysis

Tyr37-Phe-Gly-Lys-Leu-Glu-Ser-Lys-Leu-Ser

Predicted Molecular Mass

18 kDa

SDS-PAGE

18 kDa, under reducing conditions.

Product Datasheets

BT-018-GMP

Product Datasheet

COA

SDS

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.

BT-018-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS and TCEP with Trehalose.
Reconstitution	Reconstitute at 500 μg/mL in sterile water.
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. A minimum of 12 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.

Scientific Data

Bioactivity

View Larger

The bioactivity of Recombinant Human IL-18/IL-1F4 GMP Protein (Catalog # BT-018-GMP) was measured by its ability to induce IFN-gamma secretion by KG-1 human acute myelogenous leukemia cells in the presence of TNF-alpha. Three independent lots were tested for bioactivity and plotted on the same graph to show lot-to-lot consistency of GMP IL-18/IL-1F4 protein.

Bioactivity

View Larger

Equivalent bioactivity of GMP (Catalog # BT-018-GMP) and Animal-Free (BT-018-AFL) grades of Recombinant Human IL-18/IL-1F4 as measured by its ability to induce IFN-gamma secretion by KG-1 human acute myelogenous leukemia cells in the presence of TNF-alpha (orange and green, respectively).

SDS-PAGE

View Larger

2 μg/lane of Recombinant Human IL‑18/IL‑1F4 GMP Protein (Catalog # BT-018-GMP) was resolved with SDS-PAGE under reducing (R)and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing a band at 18 kDa under reducing conditions.

Data

View Larger

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM. A: Total expansion of CD56+ CD3- cells between day 0 and day 9. B: PBMCs grown in ExCellerate NK Cell Expansion Media supplemented with cytokines exhibit viability greater than 80% at day 9.

Data

View Larger

Donor PBMCs were grown using ExCellerate NK Cell Expansion Media (Catalog # CCM037) supplemented with IL-2 (Catalog # BT-002-GMP), IL-12 (Catalog # 10018-IL), IL-15 (Catalog # BT-015-GMP), IL-18 (Catalog # BT-018-GMP), and IL-21 (Catalog # BT-021-GMP) on plates coated with anti-human NKp46 (Catalog # MAB1850). PBMCs were plated to contain 1.8e4 NK cells as determined by flow cytometry on day 0. On day 5, 0.1e6 total cells were taken to seed a 6 well plate. Viability, expansion, and NK purity were analyzed on day 9, with n=5 +/- SEM for day 9; n=1 for day 0. A: Purity of NK cells at day 9. B: NK ExCellerate media preferentially expands CD56+ NK cells. Percentage of CD56+ cells in culture rose from less than 10% in both donors to more than 60% by day 9.

Reconstitution Calculator

Background: IL-18/IL-1F4

Interleukin-18 (IL-18) is a proinflammatory cytokine in the IL-1 family that exerts distinct immune effects depending on the local cytokine environment. It is expressed as a 24 kDa precursor by endothelial and epithelial cells, keratinocytes, gamma δ T cells, and phagocytes. The precursor is activated intracellularly by Caspase-1 mediated proteolysis to release the 17 kDa mature cytokine. The precursor can also be released by necrotic cells for extracellular cleavage by multiple proteases. IL-18 activation is induced by infection or tissue damage and contributes to disease pathology in chronic inflammation (1-3). IL-18 binds to the widely expressed IL-18 R alpha which recruits IL-18 R beta to form the signaling receptor complex (4, 5). Its bioactivity is negatively regulated by interactions with IL-18 binding proteins and virally encoded IL-18BP homologs (6). In the presence of IL-12 or IL-15, IL-18 enhances anti-viral Th1 immune responses by inducing IFN-gamma production and the cytolytic activity of CD8⁺ T cells and NK cells (7, 8). In the absence of IL-12 or IL-15, however, IL-18 promotes production of the Th2 cytokines IL-4 and IL-13 by CD4⁺ T cells and basophils (9, 10). In the presence of IL-1 beta or IL-23, IL-18 induces the antigen-independent production of IL-17 by gamma δ T cells and CD4⁺ T cells (11). IL-18 also promotes myeloid dendritic cell maturation and triggers neutrophil respiratory burst (12, 13). In cancer, IL-18 exhibits diverse activities including enhancing anti-tumor immunity, inhibiting or promoting angiogenesis, and promoting tumor cell metastasis (14). Mature human IL-18 shares approximately 63% amino acid sequence identity with mouse and rat IL-18 (15). Alternative splicing in human ovarian cancer generates an isoform that is resistant to Caspase-1 activation (16). A cell surface form can be expressed on M-CSF induced macrophages and released in response to bacterial endotoxin (17).

References

Dinarello, C.A. et al. (2013) Front. Immunol. 4:289.
Smith, D.E. (2011) J. Leukoc. Biol. 89:383.
Gu, Y. et al. (1997) Science 275:206.
Torigoe, K. et al. (1997) J. Biol. Chem. 272:25737.
Cheung, H. et al. (2005) J. Immunol. 174:5351.
Novick, D. et al. (1999) Immunity 10:127.
Fehniger, T.A. et al. (1999) J. Immunol. 162:4511.
Yoshimoto, T. et al. (1998) J. Immunol. 161:3400.
Yoshimoto, T. et al. (2000) Nat. Immunol. 1:132.
Kroeger, K.M. et al. (2009) J. Leukoc. Biol. 86:769.
Lalor, S.J. et al. (2011) J. Immunol. 186:5738.
Li, J. et al. (2004) Cell. Immunol. 227:103.
Elbim, C. et al. (2005) Clin. Diagn. Lab. Immunol. 12:436.
Fabbi, M. et al. (2015) J. Leukoc. Biol. 97:665.
Ushio, S. et al. (1996) J. Immunol. 156:4274.
Gaggero, A. et al. (2004) Oncogene 23:7552.
Bellora, F. et al. (2012) Eur. J. Immunol. 42:1618.

Long Name

Interleukin 18

Entrez Gene IDs

3606 (Human); 16173 (Mouse); 29197 (Rat); 397057 (Porcine); 574151 (Primate)

Alternate Names

Iboctadekin; IFN-gamma-inducing factor; IGIF; IGIFIL-1 gamma; IL18; IL-18; IL-18MGC12320; IL-1F4; IL1F4iboctadekin; IL-1g; Interferon gamma-inducing factor; interleukin 18 (interferon-gamma-inducing factor); Interleukin-1 gamma; interleukin-18

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

Additional testing and documentation requested by the customer can be arranged at an additional cost.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis and St. Paul, Minnesota USA.

R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.

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