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TNF-alpha/TNFSF2 Protein, Human

74 Cited Publications
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Based on 74 publication(s) in Google Scholar

Tumour Necrosis Factor alpha (TNF alpha) is a potent pro-inflammatory cytokine. TNF alpha binds to its receptors, mainly TNFR1 and TNFR2, and then transmits molecular signals for biological functions such as inflammation and cell death. 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. TNF alpha as a proneurogenic factor activates the SAPK/JNK Pathway and can facilitate neuronal replacement and brain repair in response to brain injury. TNF-alpha/TNFSF2 Protein, Human is a recombinant protein consisting of 157 amino acids (V77-L233) and is produced in E. coli.

For research use only. We do not sell to patients.
  • Species: Human
  • Source: E. coli
  • Storage:
    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.
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • TNF-alpha/TNFSF2 Protein, Human Product Image
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  • Biological Activity
  • Technical Parameters
  • Product Properties
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Biological Activity

Description

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 is a recombinant protein consisting of 157 amino acids (V77-L233) and is produced in E. coli.

Background

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].

In Vitro

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].

In Vivo

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].

Verified Bioactivity

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 ≤178.1 pg/mL.

MCE Validation Data

  • Purity - SDS-PAGE

    Purity - SDS-PAGE

    ≥ 95%, as determined by reducing SDS-PAGE.

  • Bioactivity - Cell-Based Assay

    Bioactivity - Cell-Based 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 62.25 pg/mL, corresponding to a specific activity is 1.61×107units/mg.

Technical Parameters

  • Species Human
  • Source E. coli
  • Tag Tag Free
  • Accession

    P01375 (V77-L233)

  • Gene ID
    7124  [NCBI]
  • Molecular Construction
    • N-term
    • TGF-α (V77-L233)
      Accession # P01375
    • C-term
  • Protein Length

    Full Length of TNF, soluble form

  • Synonyms

    TNF; Tumor Necrosis Factor (TNF Superfamily, Member 2); Prev. TNFA; Tumor Necrosis Factor-Alpha; TNF-Alpha; Tumor Necrotic Factor Alpha; TNFSF2; TNF Superfamily, Member 2; Tumor Necrosis Factor Ligand Superfamily Member 2; TNF, Macrophage-Derived; TNF-A; TNF, Monocyte-Derived; DIF; APC1 Protein; Cachectin; IMD127; TNLG1F; Tumor Necrosis Factor; Tumor Necrosis Factor Ligand 1F

  • AA Sequence

    VRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAESGQVYFGIIAL

  • Predicted Molecular Mass

    17.5 kDa

  • Molecular Weight

    Approximately 16-17 kDa, based on SDS-PAGE under reducing conditions, due to the glycosylation.

  • Purity

    ≥ 95%, as determined by reducing SDS-PAGE.

Product Properties

Appearance

Lyophilized powder

Formulation

1.Lyophilized from a 0.22 μm filtered solution of PBS.
2.Lyophilized from a 0.22 μm filtered solution of 20 mM PB, 6% Sucrose, 4% Mannitol, 0.05% Tween 80, pH 6.0.
3.Lyophilized from a 0.22 μm filtered solution of 50 mM Tris-HCl, 300 mM NaCl, pH 8.0.
4.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose, 8% mannitol and 0.02% Tween80.
5.Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4, 8% trehalose.
Please refer to the lot-specific COA for specific buffer information.
Note: For SPR assay, please replace the buffer. Primary amine components (e.g., Tris, imidazole) can affect protein-coupled chips.

Endotoxin Level

<1 EU/μg, determined by LAL method.

Reconstitution

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).

Storage & Stability

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.

Shipping

Room temperature in continental US; may vary elsewhere.

References

[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]. Zelová H, et al. TNF-α signalling and inflammation: interactions between old acquaintances. Inflamm Res. 2013 Jul;62(7):641-51. [Content Brief]

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

[4]. Ren H, et al. Matrix metalloproteinase-mediation of tumor targeting human recombinant tumor necrosis factor-α fusion protein. Mol Med Rep. 2015 Aug;12(2):2035-42. [Content Brief]

[5]. 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]

[6]. 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]

[7]. 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]

[8]. 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]

[9]. 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]

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Calculators

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The Specific Activity Calculator Equation
  • Specific Activity (Unit/mg)
  • Biological Activity (ED50)

Specific Activity (Unit/mg) = 106 ÷ Biological Activity (ED50)

Specific Activity (Unit/mg) Specific Activity (Unit/mg)
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Biological Activity (ED50) Biological Activity (ED50)
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MOQ
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100 mg

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Measured by its ability to inhibit anti-CD3-induced proliferation of stimulated CTLL-2 mouse cytotoxic T cells. The ED50 this effect is 0.8023 μg/ml, corresponding to a specific activity is 1246.42 units/mg.

Assay Types
  • 1/16

    MedChemExpress Validation
     ≥ 95%, as determined by reducing SDS-PAGE.

  • 1/16

    MedChemExpress Validation
     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 62.25 pg/mL, corresponding to a specific activity is 1.61×107units/mg.

  • 1/16

    TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2025 Aug 7:e03480. [Abstract]

    Customer Validation
     Line chart showing ACKR1 gene expression in EA.hy926 cells under different concentrations of TNF‐α (0-100 ng/ml; 48 h ).

  • 1/16

    TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 Aug;11(32):e2404171. [Abstract]

    Customer Validation
     Relative mRNA expression of (a) ICAM-1, (b) VCAM-1, (c) IL-1β, and (d) IL-10 before and after TNF-α (100 ng/mL; 4 h) stimulation.

  • 1/16

    TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Host Microbe. 2022 Oct 12;30(10):1450-1463.e8. [Abstract]

    Customer Validation
     TNF-α (50 ng/mL; 6 h); GSK2795039 (25 μM; 6 h). NETs were detected using immunofluorescent staining of DNA (DAPI, blue), dead cells and extracellular DNA (Sytox, green), histone 3 (pink), and myeloperoxidase (MPO, red). Scale bars, 50 μm.

  • 1/16

    TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Mol Biol Lett. 2022 Sep 23;27(1):79. [Abstract]

    Customer Validation
     Western blot analysis of protein expression in the indicated cells treated with TNF-α (10 ng/ml). GAPDH was used as a loading control.

  • 1/16

    TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Mol Biol Lett. 2022 Sep 23;27(1):79. [Abstract]

    Customer Validation
     TNF-α(20 ng/mL; 10 min). Confocal images of immunofluorescence staining of p65 and p-p65 in the indicated cells.

  • TNF-alpha/TNFSF2 Protein, Human Product Image
    1/16
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • Experimental Validation Results for TNF-alpha/TNFSF2 Protein, Human
  • TNF-alpha/TNFSF2 Protein, Human Product Image

  • TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2025 Aug 7:e03480. [Abstract]

    Line chart showing ACKR1 gene expression in EA.hy926 cells under different concentrations of TNF‐α (0-100 ng/ml; 48 h ).

  • TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2024 Aug;11(32):e2404171. [Abstract]

    Relative mRNA expression of (a) ICAM-1, (b) VCAM-1, (c) IL-1β, and (d) IL-10 before and after TNF-α (100 ng/mL; 4 h) stimulation.

  • TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Host Microbe. 2022 Oct 12;30(10):1450-1463.e8. [Abstract]

    TNF-α (50 ng/mL; 6 h); GSK2795039 (25 μM; 6 h). NETs were detected using immunofluorescent staining of DNA (DAPI, blue), dead cells and extracellular DNA (Sytox, green), histone 3 (pink), and myeloperoxidase (MPO, red). Scale bars, 50 μm.

  • TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Mol Biol Lett. 2022 Sep 23;27(1):79. [Abstract]

    Western blot analysis of protein expression in the indicated cells treated with TNF-α (10 ng/ml). GAPDH was used as a loading control.

  • TNF-alpha/TNFSF2 Protein, Human purchased from MedChemExpress. Usage Cited in: Cell Mol Biol Lett. 2022 Sep 23;27(1):79. [Abstract]

    TNF-α(20 ng/mL; 10 min). Confocal images of immunofluorescence staining of p65 and p-p65 in the indicated cells.