Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF

Recombinant human TNF-alpha (Catalog # 10291-TA) has a molecular weight (MW) of 52.3 kDa as analyzed by SEC-MALS, suggesting that this protein is a homotrimer. MW may differ from predicted MW due to post-translational modifications (PTMs) present (i.e. Glycosylation).

Recombinant Human TNF‑ alpha (HEK293-expressed) (Catalog # 10291-TA) induces cytotoxicity in the L‑929 mouse fibroblast cell line in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 20‑100 pg/mL.

2 μg/lane of Recombinant Human TNF-alpha (HEK293-expressed) (Catalog # 10291-TA) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 14-18 kDa.

• Reactivity: Hu
• Applications: Bioactivity
• Format: Carrier-Free

Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF Summary

• Additional Information: Analyzed by SEC-MALS
• Details of Functionality: 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 20-100 pg/mL.
• Source: Human embryonic kidney cell, HEK293-derived human TNF-alpha protein
  Val77-Leu233
• Accession #: P01375
• N-terminal Sequence: Val77
• Protein/Peptide Type: Recombinant Proteins
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Endotoxin Note: <0.10 EU per 1 μg of the protein by the LAL method.

Applications/Dilutions

• Dilutions:
  • Bioactivity
• Theoretical MW: 17 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• SDS-PAGE: 14-18 kDa, under reducing conditions

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose.
• Purity: >95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
• Reconstitution Instructions: Reconstitute at 500 μg/mL in PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human TNF-alpha (HEK293-expressed) Protein, CF

• 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

Background

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).

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

Publications for TNF-alpha (10291-TA)(11)

Publications using 10291-TA

• Sun, Y;Maggs, L;Panda, A;Wright, SJ;Cicerchia, AM;Jenney, A;Perricone, MD;Mills, CE;Cattaneo, G;Ventin, M;Chen, F;Rasmussen, MQ;Miranda, A;Revach, OY;Fang, J;Fu, A;Bowling, PJ;Sharova, T;Lawless, A;Sorger, PK;Bardeesy, N;Wang, X;Flaherty, KT;Boland, GM;Mehta, A;Sade-Feldman, M;Ferrone, CR;Jenkins, RW; TBK1 Targeting Is Identified as a Therapeutic Strategy to Enhance CAR T-Cell Efficacy Using Patient-Derived Organotypic Tumor Spheroids Cancer immunology research 2025-01-09 [PMID: 39785827] (Cell Culture, Human)
• Majocchi, S;Lloveras, P;Nouveau, L;Legrand, M;Viandier, A;Malinge, P;Charreton, M;Raymond, C;Pace, E;Millard, B;Svensson, L;Kelpšas, V;Anceriz, N;Salgado-Pires, S;Daubeuf, B;Magistrelli, G;Gueneau, F;Moine, V;Masternak, K;Shang, L;Fischer, N;Ferlin, W; NI-3201 Is a Bispecific Antibody Mediating PD-L1-Dependent CD28 Co-stimulation on T Cells for Enhanced Tumor Control Cancer Immunology Research 2025-01-06 [PMID: 39760515] (Bioassay, Human)
• Li, Y;Li, TY;Qiao, Q;Zhang, MT;Tong, MX;Xu, LF;Zhang, ZB; Polymeric immunoglobulin receptor promotes Th2 immune response in the liver by increasing cholangiocytes derived IL-33: a diagnostic and therapeutic biomarker of biliary atresia EBioMedicine 2024-09-16 [PMID: 39288533] (Cell Culture, Human)
• Hayderi, A;Zegeye, MM;Meydan, S;Sirsjö, A;Kumawat, AK;Ljungberg, LU; TNF Induces Laminin-332-Encoding Genes in Endothelial Cells and Laminin-332 Promotes an Atherogenic Endothelial Phenotype International journal of molecular sciences 2024-08-09 [PMID: 39201392] (Bioassay, Human)
• Ulrich, A;Wu, Y;Draisma, H;Wharton, J;Swietlik, EM;Cebola, I;Vasilaki, E;Balkhiyarova, Z;Jarvelin, MR;Auvinen, J;Herzig, KH;Coghlan, JG;Lordan, J;Church, C;Howard, LS;Pepke-Zaba, J;Toshner, M;Wort, SJ;Kiely, DG;Condliffe, R;Lawrie, A;Gräf, S;Morrell, NW;Wilkins, MR;Prokopenko, I;Rhodes, CJ; Blood DNA methylation profiling identifies cathepsin Z dysregulation in pulmonary arterial hypertension Nature communications 2024-01-06 [PMID: 38184627] (Bioassay, Human)
• Mijit, M;Kpenu, E;Chowdhury, NN;Gampala, S;Wireman, R;Liu, S;Babb, O;Georgiadis, MM;Wan, J;Fishel, ML;Kelley, MR; In vitro and In vivo evidence demonstrating chronic absence of Ref-1 Cysteine 65 impacts Ref-1 folding configuration, redox signaling, proliferation and metastasis in pancreatic cancer Redox biology 2023-12-01 [PMID: 38056311] (Bioassay, Human)
• Barilli, A;Recchia Luciani, G;Visigalli, R;Sala, R;Soli, M;Dall'Asta, V;Rotoli, BM; Cytokine-Induced iNOS in A549 Alveolar Epithelial Cells: A Potential Role in COVID-19 Lung Pathology Biomedicines 2023-10-03 [PMID: 37893073] (Bioassay, Human)
• Lassé, M;El Saghir, J;Berthier, CC;Eddy, S;Fischer, M;Laufer, SD;Kylies, D;Hutzfeldt, A;Bonin, LL;Dumoulin, B;Menon, R;Vega-Warner, V;Eichinger, F;Alakwaa, F;Fermin, D;Billing, AM;Minakawa, A;McCown, PJ;Rose, MP;Godfrey, B;Meister, E;Wiech, T;Noriega, M;Chrysopoulou, M;Brandts, P;Ju, W;Reinhard, L;Hoxha, E;Grahammer, F;Lindenmeyer, MT;Huber, TB;Schlüter, H;Thiel, S;Mariani, LH;Puelles, VG;Braun, F;Kretzler, M;Demir, F;Harder, JL;Rinschen, MM; An integrated organoid omics map extends modeling potential of kidney disease Nature communications 2023-08-14 [PMID: 37580326] (Bioassay, Human)
• Isozaki, H;Sakhtemani, R;Abbasi, A;Nikpour, N;Stanzione, M;Oh, S;Langenbucher, A;Monroe, S;Su, W;Cabanos, HF;Siddiqui, FM;Phan, N;Jalili, P;Timonina, D;Bilton, S;Gomez-Caraballo, M;Archibald, HL;Nangia, V;Dionne, K;Riley, A;Lawlor, M;Banwait, MK;Cobb, RG;Zou, L;Dyson, NJ;Ott, CJ;Benes, C;Getz, G;Chan, CS;Shaw, AT;Gainor, JF;Lin, JJ;Sequist, LV;Piotrowska, Z;Yeap, BY;Engelman, JA;Lee, JJ;Maruvka, YE;Buisson, R;Lawrence, MS;Hata, AN; Therapy-induced APOBEC3A drives evolution of persistent cancer cells Nature 2023-07-05 [PMID: 37407818] (Immunofluorescence, Human)
• SW Wang, Q Zhang, D Lu, YC Fang, XC Yan, J Chen, ZK Xia, QT Yuan, LH Chen, YM Zhang, FJ Nan, X Xie GPR84 regulates pulmonary inflammation by modulating neutrophil functions Acta pharmacologica Sinica, 2023-04-04;0(0):1-11. 2023-04-04 [PMID: 37016043] (Bioassay, Mouse)
• S Salva, I Tiscornia, F Gutiérrez, S Alvarez, M Bollati-Fo Lactobacillus rhamnosus postbiotic-induced immunomodulation as safer alternative to the use of live bacteria Cytokine, 2021-07-10;146(0):155631. 2021-07-10 [PMID: 34252871] (Bioassay, Human)

FAQs for TNF-alpha (10291-TA). (Showing 1 - 4 of 4 FAQs).

1. I need to know about TNF-alpha antibody kits for mice.
   • We currently have two TNFalpha ELISA kits specific for mouse: catalog numbers KA0257 and NBP1-92670.
2. I would like to ask you for help. I need an antibody for Elisa to detect human TNF alpha in a supernatant from a cell culture (meaning supernatant after centrifugation of collected cell suspension from a plate well). Which of your antibodies against human tnf alpha would be suitable? I would like to buy only the primary antibody.
   • I would recommend catalog number NBP1-67821 or NB600-587; however, a full list of our anti-human TNF alpha antibodies suitable for use in ELISA can be found using this link.
3. I am interested in a TNF alpha antibody, cross reactive for human, rat and mouse (host species: rabbit). Could your product NBP1-19532 be used in western blotting applications? Or do you have a similar product in your catalog which could fit with my request?
   • The antibody you mention, NBP1-19532, has not yet been validated in Western blot. I would instead recommend either NBP1-67821 or NB600-587. These are both rabbit polyclonal antibodies that cross-reacts with human, mouse and rat and have been used in Western blotting.
4. Can your TNF-alpha products be used to treat TBI victims and therefore avoid the perispinal injections with Enbrel?
   • I am very sorry, but all of our products are for scientific research use only, and none are intended or approved for use in humans.

Bioinformatics

• Uniprot:
  • P01375()