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0032 (0)16 41 44 07
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Česká republika Praha
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0032 (0)16 41 44 07
Recombinant Human Tumor Necrosis Factor alpha (TNFa)
(Cat. No.: C008)
TNF-alpha is a homotrimer with a subunit molecular mass of 17 kDa and that it plays a major role in growth regulation, differentiation, inflammation, viral replication, tumorigenesis, and autoimmune diseases; and in viral, bacterial, fungal, and parasitic infections. Besides inducing hemorrhagic necrosis of tumors, TNF was found to be involved in tumorigenesis, tumor metastasis, viral replication, septic shock, fever, inflammation, and autoimmune diseases including Crohn’s disease, and rheumatoid arthritis as well as graft-versus-host disease.
Tumor necrosis factor alpha-1a or TNFalpha-1a is a non-glycosylated cytokine produced from E. coli using rDNA technology. The protein consists of three identical polypeptide chains of 158 amino acids combined to form a compact, bell-shaped homotrimer. The individual subunits have a relative molecular mass each of 17,484 Daltons. TNF alpha-1a is a potent lymphoid factor that exerts cytotoxic effects on a wide range of tumor cells and certain other target cells.
Biological activity: The ED50 as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D is less then 0.03ng/ml, corresponding to a Specific Activity of 3.0 x 107 IU/mg.
Purity: Greater than 95% as determined by
(a) Analysis by SEC-HPLC.
(b) Analysis by reducing and non-reducing SDS-PAGE Silver Stained gel.
Molecular weight: 17.3 KD+/-10% determined by reduced SDS-PAGE.
Isoelectric Point: the main zone between 4.0~5.0 analysis by IEF.
UV scan: the maximal absorption wave is 275+/-3nm.
Amino-Acid Sequence: The sequence of the first fifteen N-terminal amino acids was determined and was found to be Met-Val-Arg-Ser-Ser-Ser-Arg-Thr-Pro-Ser-Asp-Lys-Pro-Val- Ala
Residual DNA: less than 10ng/mg analysis by solid phase blot.
Residual host cell protein: less than 0.1% analysis by ELISA.
Endotoxin: Less than 0.01ng/µg (0.01IEU/µg) determined by LAL test.
Formulation: Lyophilized from a 0.2 µm filtered solution in PBS.
Storage: Lyophilized samples are stable for up to twelve months from date of receipt at -20oC to -70oC.
Please avoid freeze-thaw cycles.
Reconstitution: It is recommended to reconstitute the lyophilized TNF alpha-1a in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
1: Mol Nutr Food Res 2009 Jan;
Silibinin protects OTA-mediated TNF-alpha release from perfused rat livers and isolated rat Kupffer cells.
[Abstract] We studied the inhibitory effect of silibinin on ochratoxin A (OTA) and LPS-mediated tumor necrosis factor alpha (TNF-alpha) release and the leakage of cytotoxic markers glutamate dehydrogenase (GLDH) and lactate dehydrogenase (LDH), from isolated blood-free perfused rat livers, and from isolated pure rat Kupffer cells. In the recirculation perfusion model at the end point 90 min, 2.5 mumol/L OTA released 2600 pg/mL TNF-alpha without effects on liver vitality. LPS at 0.1 mug/mL induced 3000 pg TNF-alpha/mL with slight leakage of GLDH and LDH. Under similar experimental conditions, the addition of silibinin 10 min prior to OTA and LPS showed dose-dependent protection against OTA or LPS-induced hepatic TNF-alpha release. High-dose of silibinin (12.5 mug/mL) also
completely restored GLDH and LDH levels in the perfusate. Pretreatment of isolated Kupffer cells with 0.02, 0.1, 0.5, 2.5, and 12.5 mug silibinin/mL 30 min prior to OTA reduced OTA-induced TNF-alpha levels to 90, 70, 25, 25, and 25% at 4 h, respectively, and abrogated any TNF-alpha release at 24 h. Similarly, in the presence of silibinin LPS-induced TNF-alpha levels decreased at 4 h to 71, 57, 18, 22, and 18%, respectively. However, after 24 h of LPS exposition the protection by silibinin vanished and TNF-alpha partially recurred into the incubation medium under LPS. In summary, silibinin had hepatoprotective effects against OTA- or LPS-mediated TNF-alpha release and also reduced the cytotoxicity of both toxins. Isolated Kupffer cells were even more sensitive to the protective effect than
perfused livers and responded to very low concentrations of silibinin with a strong inhibition of toxins-mediated TNF-alpha release.
2: PLoS ONE 2009 ;Vol 4(1)
Pleural tuberculosis in patients with early HIV infection is associated with increased TNF-alpha expression and necrosis in granulomas.
[Abstract] Although granulomas may be an essential host response against persistent antigens, they are also associated with immunopathology. We investigated whether HIV co-infection affects histopathological appearance and cytokine profiles of pleural granulomas in patients with active pleural tuberculosis (TB). Granulomas were investigated in pleural biopsies from HIV positive and negative TB pleuritis patients. Granulomas were characterised as necrotic or non-necrotic, graded histologically and investigated for the mRNA expression of IL-12, IFN-gamma, TNF-alpha and IL-4 by in situ hybridisation. In all TB patients a mixed Th1/Th2 profile was noted. Necrotic granulomas were more evident in HIV positive patients with a clear association between TNF-alpha and necrosis.
This study demonstrates immune dysregulation which may include TNF-alpha-mediated immunopathology at the site of disease in HIV infected pleural TB patients.
3: Transplantation 2009
Intragraft TNF receptor signaling contributes to activation of innate and adaptive immunity in a renal allograft model.
[Abstract] BACKGROUND: Increased levels of tumor necrosis factor (TNF) are a risk factor for allograft rejection. In vitro studies have shown that binding of TNF to its receptor activates signaling cascades that induce expression of many genes involved in inflammation. The role of intragraft TNF receptor (TNFR) signaling in activation of gene expression in allografts has not been studied. METHODS: Gene expression profiling and quantitative real-time polymerase chain reaction analysis were used to investigate the role of TNFR signaling in the early intragraft activation of cellular gene expression in renal allografts at 2 days posttransplant. RESULTS: The TNFRs play a critical role in activating intragraft expression of transcription factors controlling innate and
adaptive immunity and stress responses (interferon regulatory factor [IRF]1, IRF 8, Isgf3g, and ATF3) of cytokines and receptors mediating inflammation (TNF, interleukin [IL]-6, interferon-gamma, oncostatin M receptor [OMCR], toll-like receptor [TLR]2, and IL-2Rgamma), of chemokines and adhesion molecules that recruit inflammatory cells (Cxcl9, Cxcl11, E-selectin, and intracellular adhesion molecule [ICAM]-1), of genes involved in costimulation of T cells and processing and presentation of antigens (H2-DMb, Psmb8, and CD40), and genes that mediate the response to interferons. In addition to its proinflammatory role, TNFR signaling induces expression of SOCS3, a negative regulator of IL-6 and OSMR signaling and Nfkbie, and a negative regulator of TNFR signal transduction. CONCLUSIONS: These
studies illustrate the pleiotropic effect of TNF in both activation and down-modulation of the immune response and the complex interactions between the TNFRs and other cytokine signaling pathways in the early allograft response.
4: J Nutr Sci Vitaminol (Tokyo) 2008
Effect of dietary soy protein on tumor necrosis factor productivity in macrophages from nephritic and hepatoma-bearing rats.
[Abstract] The present study investigated the effect of dietary soy protein isolate (SPI) on tumor necrosis factor-alpha (TNF) productivity in peritoneal macrophages from nephritic and hepatoma-bearing rats. Dietary SPI significantly inhibited the elevated production of TNF by lipopolysaccharide-stimulated macrophages in nephritic and hepatoma-bearing rats compared with dietary casein, while it exerted no influence on the TNF productivity in normal rats. Removal of the minor components contained in SPI by ethanol extraction could significantly or partially restore the reduced TNF production caused by SPI in nephritic and hepatoma-bearing rats, respectively. These results suggest that dietary SPI could suppress the enhanced productivity of TNF associated with the
progression of nephritis and hepatoma, and some factors existing in the ethanol extract of SPI are suggested to be involved in suppressing TNF productivity by macrophages.
Protein Accession No.:
M V R S S S R T P S D K P V A H V V A N P Q A E G Q L Q W L N R R A N A L L A N G V E L R D N Q L V V P S E G L Y L I Y S Q V L F K G Q G C P S T H V L L T H T I S R I A V S Y Q T K V N L L S A I K S P C Q R E T P E G A E A K P W Y E P I Y L G G V F Q L E K G D R L S A E I N R P D Y L D F A E S G Q V Y F G I I A L
*NOTE: ALL PRODUCTS ARE FOR RESEARCH USE ONLY