Recombinant Human TNF-alpha, Animal-Free Protein

Name: Recombinant Human TNF-alpha, Animal-Free Protein
Brand: R&D Systems
SKU: AFL210
Rating: 5 (1 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # AFL210

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Intended for preclinical researchers who may transition to GMP TNF-alpha for their clinical work

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Learn more about Animal-Free Recombinant Proteins

Source:	E. coli
Accession #:	P01375.1
Applications:	Bioactivity

View Full Technical Details for Recombinant Human TNF-alpha, Animal-Free Protein

Product Specifications for Recombinant Human TNF-alpha, Animal-Free Protein

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.
Purity	>97%, 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	Met & Val77
Predicted Molecular Mass	17 kDa
SDS-PAGE	17 kDa, reducing conditions
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).

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.

AFL210

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS.
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.

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: 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).

Item	Value
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, TNF-A, TNF-alpha, TNF-alphacachectin, TNFA, TNFATNF, macrophage-derived, TNFG1F, TNFSF1A, TNFSF2, TNFSF2TNF superfamily, member 2, TNFalpha, Tumor necrosis factor ligand superfamily member 2, tumor necrosis factor, tumor necrosis factor (TNF superfamily, member 2), tumor necrosis factor alpha, tumor necrosis factor-alpha

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

Citations for Recombinant Human TNF-alpha, Animal-Free Protein

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Customer Reviews for Recombinant Human TNF-alpha, Animal-Free Protein (1)

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Recombinant Human TNF-alpha, Animal-Free Protein

Name: Martin Torres

Application: In vitro bioactivity in cell culture

Reason for Rating: TNF-a protein was used successfully as a control of mice-derived pre-adipocytes activation during their maturation to adipocytes.

Verified Customer | Posted 10/22/2021

C57BL/6J-derived pre-adipocytes were extracted from adipose tissue. After culturing for 3 weeks, cells were stimulated with TNF-alpha as pro-inflammatory control, while the other cells were treated with a Moringa oleifera extract (10, 50, and 100 micrograms/mL). The quantification of Reactive oxygen species in the cells showed an adequate impact of TNF-alpha inducing inflammation and ROS activation. ROS were measured by fluorescence in relative fluorescence units (RFU).

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Recombinant Human TNF-alpha, Animal-Free Protein

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Product Specifications for Recombinant Human TNF-alpha, Animal-Free Protein

Formulation, Preparation and Storage

Carrier Free

Reconstitution Calculator

Background: TNF-alpha

Manufacturing Specifications

Citations for Recombinant Human TNF-alpha, Animal-Free Protein

Customer Reviews for Recombinant Human TNF-alpha, Animal-Free Protein (1)

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Recombinant Human TNF-alpha, Animal-Free Protein

Product Documents for Recombinant Human TNF-alpha, Animal-Free Protein

Product Datasheets

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