Recombinant Human TNF-alpha GMP Protein, CF

Catalog #: 210-GMP
3 Citations Datasheet / COA / SDS

Animal-Free

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210-GMP-100
210-GMP-02M

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Recombinant Human TNF-alpha GMP Protein Bioactivity

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Product Details

Citations (3)

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

Recombinant Human TNF-alpha GMP Protein, CF Summary

Product Specifications

Purity

>97%, by SDS-PAGE with silver staining

Endotoxin Level

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

Activity

Measured in a cytotoxicity assay using L‑929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach. Clemens, M.J. et al. (eds): IRL Press. 221. The ED₅₀ for this effect is 25-100 pg/mL.
The specific activity of recombinant human TNF-alpha GMP is >4.3 x 10⁷ IU/mg, which is calibrated against the human TNF-alpha WHO International Standard (NIBSC code: 12/154).

Source

E. coli-derived human TNF-alpha protein
Val77-Leu233, with and without an N-terminal Met
Produced using non-animal reagents in an animal-free laboratory.
Manufactured and tested under cGMP guidelines.

Accession #

P01375.1

N-terminal Sequence
Analysis

Met-Val₇₇-Arg-Ser-Ser-Ser-Arg-Thr-Pro-Ser
Val₇₇-Arg-Ser-Ser-Ser-Arg-Thr-Pro-Ser-Asp

Predicted Molecular Mass

17 kDa

SDS-PAGE

17 kDa, reducing conditions

Product Datasheets

210-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.

210-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 0.1-1 mg/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. 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

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GMP-grade Recombinant Human TNF-alpha (Catalog # 210-GMP) induces cytotoxicity in the L-929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED₅₀ is 25-100 pg/mL.

SDS-PAGE

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1 μg/lane of GMP-grade Recombinant Human TNF-alpha (Catalog # 210-GMP) was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a single band at 17 kDa.

Mass Spectrometry

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MALDI-TOF analysis of GMP-grade Recombinant Human TNF-alpha (Catalog # 210‑GMP). The major peak at 17348 corresponds to the calculated molecular mass without an N-terminal Met, 17353 Da. The minor peak at 17480 corresponds to the calculated molecular mass with an N-terminal Met, 17484 Da. The peak at 17565 is a matrix-associated artifact of the MALDI-TOF.

Reconstitution Calculator

Background: TNF-alpha

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (1, 2). Human TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 177 aa extracellular domain (ECD) (3). Within the ECD, human TNF-alpha shares 97% aa sequence identity with rhesus and 71%-92% with bovine, canine, cotton rat, equine, feline, mouse, porcine, and rat TNF-alpha. TNF-alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells (1, 2). TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface (4). Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I (2, 5). Shedding of membrane bound TNF-alpha by TACE/ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer of the TNF-alpha extracellular domain (6-8). TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (9, 10) and the hematopoietic cell-restricted 80 kDa TNF RII (11, 12), both of which are also expressed as homotrimers (1, 2, 13). Both type I and type II receptors bind TNF-alpha with comparable affinity (14), although only TNF RI contains a cytoplasmic death domain which triggers the activation of apoptosis. Soluble forms of both types of receptors are released and can neutralize the biological activity of TNF-alpha (15).

References

Zelova, H. and J. Hosek (2013) Inflamm. Res. 62:641.
Juhasz, K. et al. (2013) Expert Rev. Clin. Immunol. 9:335.
Pennica, D. et al. (1984) Nature 312:724.
Tang, P. et al. (1996) Biochemistry 35:8216.
Perez, C. et al. (1990) Cell 63:251.
Black, R.A. et al. (1997) Nature 385:729.
Moss, M.L. et al. (1997) Nature 385:733.
Gearing, A.J.H. et al. (1994) Nature 370:555.
Schall, T.J. et al. (1990) Cell 61:361.
Loetscher, H. et al. (1990) Cell 61:351.
Dembic, Z. et al. (1990) Cytokine 2:231.
Smith, C.A. et al. (1990) Science 248:1019.
Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324.
Pinckard, J.K. et al. (1997) J. Biol. Chem. 272:10784.
Engelmann, H. et al. (1990) J. Biol. Chem. 265:1531.

Long Name

Tumor Necrosis Factor alpha

Entrez Gene IDs

7124 (Human); 21926 (Mouse); 24835 (Rat); 397086 (Porcine); 280943 (Bovine); 403922 (Canine); 102139631 (Cynomolgus Monkey); 100033834 (Equine); 493755 (Feline); 100009088 (Rabbit)

Alternate Names

APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha; TNF-alpha; TNF-alphacachectin; TNFATNF, macrophage-derived; TNFG1F; TNFSF1A; TNFSF2; TNFSF2TNF superfamily, member 2; tumor necrosis factor (TNF superfamily, member 2); tumor necrosis factor alpha; Tumor necrosis factor ligand superfamily member 2; tumor necrosis factor; tumor necrosis factor-alpha

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

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.

Please read our complete Animal-Free Statement

Product Specific Notices

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

Related Research Areas

Citations for Recombinant Human TNF-alpha GMP Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

3 Citations: Showing 1 - 3
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Autologous HIV-specific T cell therapy targeting conserved epitopes is well-tolerated in six adults with HIV: an open-label, single-arm phase 1 study
Authors: Sohai, DK;Keller, MD;Hanley, PJ;Hoq, F;Kukadiya, D;Datar, A;Reynolds, E;Copertino, DC;Lazarski, C;McCann, CD;Tanna, J;Shibli, A;Lang, H;Zhang, A;Chansky, PA;Motta, C;Huynh, TT;Dwyer, B;Wilson, A;Lynch, R;Mota, TM;Conce Alberto, WD;Brumme, ZL;Kinloch, NN;Cruz, CRY;MacLaren Ehui, L;Henn, S;Brad Jones, R;Bollard, CM;
Nature communications
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
Opposing roles of hematopoietic-specific small GTPase Rac2 and the guanine nucleotide exchange factor Vav1 in osteoclast differentiation
Authors: IS Kang, JS Jang, C Kim
Sci Rep, 2020-04-27;10(1):7024.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay
The IAP Antagonist SM-164 Eliminates Triple-Negative Breast Cancer Metastasis to Bone and Lung in Mice
Authors: W Lei, R Duan, J Li, X Liu, A Huston, BF Boyce, Z Yao
Sci Rep, 2020-04-24;10(1):7004.
Species: Human
Sample Types: Whole Cells
Applications: Bioassay