TNF alpha/TNFSF2 protein, Human (His)

Tumour Necrosis Factor alpha (TNF alpha) is a potent pro-inflammatory cytokine^[1]. TNF alpha binds to its receptors, mainly TNFR1 and TNFR2, and then transmits molecular signals for biological functions such as inflammation and cell death^[2]. TNF alpha stimulates NF-κB pathway via TNFR2 promotes cancer growth, invasion, and metastasis. Anti-TNF-α MAb significantly suppresses the tumor development in colitis-associated cancer (CAC) mouse^[3]. TNF alpha as a proneurogenic factor activates the SAPK/JNK Pathway and can facilitate neuronal replacement and brain repair in response to brain injury^[4]. TNF alpha/TNFSF2 protein, Human (His) is a recombinant protein with a N-Terminal His label, It consists of 157 amino acids (V77-L233) and is produced in CHO cells.

TNF alpha is produced by various types of cells including macrophages, monocytes, neutrophils, T cells, and NK-cells^[2].
The amino acid sequence of human TNF alpha protein has low homology between mouse, rat, bovine, cynomolgus TNF alpha protein. While, human TNF alpha shares 94.85% aa sequence identity with cynomolgus TNF alpha protein, mouse TNF alpha shares 94.47% aa sequence identity with rat TNF alpha protein.
TNF alpha exists in two forms; a type II transmembrane protein (tmTNF-α) and a mature soluble protein (sTNF-α). TNF-α binds to its receptors, mainly TNFR1 and TNFR2, and then transmits molecular signals for biological functions such as inflammation and cell death. Both sTNF-α and tmTNF-α activate TNFR1, and process a death domain (DD) that interacts with the TNFR1-associated death domain (TRADD) adaptor protein. The TNFR2 signaling pathway is mainly activated by tmTNF-α. TNFR1 signaling tends to be pro-inflammatory and apoptotic. TNFR2 results in NF-κB and MAPKs and AKT activation, TNFR2 activation is associated with homeostatic bioactivities such as tissue regeneration, cell proliferation, and cell survival, as well as host defense and inflammation^[1].
TNF-alpha is critical for normal immune response, abnormal secretion TNF alpha activates synovial fibroblasts, keratinocytes, osteoclasts, induces rheumatoid arthritis, inflammatory bowel disease, psoriatic arthritis (PsA), and noninfectious uveitis (NIU)^[3]. TNF alpha positively regulates endogenous TNF-α expression levels independently of Pgp efflux activity, induces IHF cells proliferation^[4]. TNF alpha in tissues may promote cancer growth, invasion, and metastasis. Besides, TNF alpha stimulates NF-κB pathway via TNFR2 and anti-TNF-α MAb significantly suppresses the tumor development in colitis-associated cancer (CAC) mouse^[5]. TNF alpha as a proneurogenic factor activates the SAPK/JNK pathway and can facilitate neuronal replacement and brain repair in response to brain injury^[6].

TNF alpha (human) (0, 10, 15, 20, 30, 50 ng/mL; 24, 48 h) reduces KB-C1 cells viability at 30 ng/mL after 48 h, reduces pro-caspase-3, cleaved caspase-3 expression in KB-C1 and KB-3-1 cells^[4].
TNF alpha (human) (10, 15 ng/mL; 24 h) significantly increases the mRNA levels of Pgp (ABCB1) in KB-3-1 cells, and promotes expressive reduction on total Pgp protein levels and a slightly decreases in cell surface-Pgp in KB-C1 cells, and stimulates IHF cells proliferation, probably via ERK activation^[4].

TNF alpha (human) (100 µg; osmium pump in back; daily for 7 days) significantly increases in the number of inflammatory cells and the degree of synovial inflammation is significantly exacerbated in twenty-four SCID-HuRAg mice^[7].

1. Measured in a cytotoxicity assay using L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED₅₀ for this effect is < 0.13 ng/mL.
2. Loaded Brivekimig (HY-P990907) on AHC2 biosensor, can bind TNF alpha protein, Human (His) with an affinity constant of <1.0E-12 M as determined in BLI assay.

• 
  Measured in a cytotoxicity assay using L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED₅₀ for this effect is 0.4678 pg/mL, corresponding to a specific activity is 2.137×10⁹ units/mg.
• 
  Loaded Brivekimig (HY-P990907) on AHC2 biosensor, can bind TNF alpha protein, Human (His) with an affinity constant of <1.0E-12 M as determined in BLI assay.

Human

E. coli

N-6*His

P01375 (V77-L233)

7124  [NCBI]

Full Length of TNF soluble form

VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL

Approximately 18 kDa, based on SDS-PAGE under reducing conditions.

• 
  ≥ 90%, as determined by reducing SDS-PAGE.

Lyophilized powder

1.Lyophilized from a 0.22 μm filtered solution of PBS, 300 mM NaCl, pH 7.4.
2.Lyophilized from a 0.22 μm filtered solution of PBS.
3.Lyophilized from a 0.22 μm filtered solution of PBS, pH 6.5, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.

• [1]. Horiuchi T, et al. Transmembrane TNF-alpha: structure, function and interaction with anti-TNF agents. Rheumatology (Oxford). 2010 Jul;49(7):1215-28.  [Content Brief]

  [2]. El-Tahan RR, et al. TNF-α gene polymorphisms and expression. Springerplus. 2016 Sep 7;5(1):1508.  [Content Brief]

  [3]. Jang DI, et al. The Role of Tumor Necrosis Factor Alpha (TNF-α) in Autoimmune Disease and Current TNF-α Inhibitors in Therapeutics. Int J Mol Sci. 2021 Mar 8;22(5):2719.  [Content Brief]

  [4]. Berguetti T, et al. TNF-α Modulates P-Glycoprotein Expression and Contributes to Cellular Proliferation via Extracellular Vesicles. Cells. 2019 May 24;8(5):500.  [Content Brief]

  [5]. Onizawa M, et al. Signaling pathway via TNF-alpha/NF-kappaB in intestinal epithelial cells may be directly involved in colitis-associated carcinogenesis. Am J Physiol Gastrointest Liver Physiol. 2009 Apr;296(4):G850-9.  [Content Brief]

  [6]. Bernardino L, et al. Tumor necrosis factor-alpha modulates survival, proliferation, and neuronal differentiation in neonatal subventricular zone cell cultures. Stem Cells. 2008 Sep;26(9):2361-71.  [Content Brief]

  [7]. Matsuno H, et al. The role of TNF-alpha in the pathogenesis of inflammation and joint destruction in rheumatoid arthritis (RA): a study using a human RA/SCID mouse chimera. Rheumatology (Oxford). 2002 Mar;41(3):329-37.  [Content Brief]