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Recombinant Tumor Necrosis Factor alpha (TNF alpha), human

Catalogue Number CYT-223
Synonyms TNF-alpha, Tumor necrosis factor ligand superfamily member 2, TNF-a, Cachectin, DIF, TNFA, TNFSF2.
Introduction Tumor necrosis factor is a cytokine involved in systemic inflammation and is a member of a group of cytokines that all stimulate the acute phase reaction. TNF is mainly secreted by macrophages.
TNF causes apoptotic cell death, cellular proliferation, differentiation, inflammation, tumorigenesis and viral replication, TNF is also involved in lipid metabolism, and coagulation. TNF's primary role is in the regulation of immune cells.
Dysregulation and, in particular, overproduction of TNF have been implicated in a variety of human diseases- autoimmune diseases, insulin resistance, and cancer.
Description Tumor Necrosis Factor-a Human Recombinant produced in E.Coli is a single, non-glycosylated, polypeptide chain containing 158 amino acids (157 a.a. of the mature human TNF-alpha and an N-terminal methionine) and having a molecular mass of 17.5kDa.
The TNF-alpha is purified by standard chromatographic techniques.
Source Escherichia Coli.
Physical Appearance Sterile Filtered White lyophilized (freeze-dried) powder.
Formulation 1mg of TNF-alpha Human contain 20mM PB, pH-7.2, and 100mM NaCl.
Solubility It is recommended to reconstitute the lyophilized Tumor Necrosis Factor-alpha in sterile 18MΩ-cm H2O not less than 100µg/ml, which can then be further diluted to other aqueous solutions.
Stability Lyophilized Tumor Necrosis Factor-a although stable at room temperature for 3 weeks, should be stored desiccated below -18°C. Upon reconstitution TNF-a should be stored at 4°C between 2-7 days and for future use below -18°C.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please prevent freeze-thaw cycles.
Purity Greater than 95.0% as determined by:
(a) Analysis by RP-HPLC.
(b) Analysis by SDS-PAGE.
Amino acid sequence MVRSSSRTPS DKPVAHVVAN PQAEGQLQWL NRRANALLAN GVELRDNQLV VPSEGLYLIY SQVLFKGQGC PSTHVLLTHT ISRIAVSYQT KVNLLSAIKS PCQRETPEGA E AKPWYEPIY LGGVFQLEKG DRLSAEINRP DYLDFAESGQ VYFGIIAL.
Biological Activity The Specific Activity is >5.0×107 IU/mg as determined by the cytolysis of murine L929 cells in the presence of Actinomycin D.
Protein content Protein quantitation was carried out by two independent methods
1. UV spectroscopy at 280 nm using the absorbency value of 1.234 as the extinction coefficient for a 0.1% (1mg/ml) solution. This value is calculated by the PC GENE computer analysis program of protein sequences (IntelliGenetics).

2. Analysis by RP-HPLC, using a calibrated solution of TNF-a as a Reference Standard.
References

1.Title: Radiation-triggered Tumor Necrosis Factor (TNF) ?-NF?B Cross-signaling Favors Survival Advantage in Human Neuroblastoma Cells*
Publication:First Published on April 28, 2011, doi: 10.1074/jbc.M110.193755 June 17, 2011 The Journal of Biological Chemistry, 286, 21588-21600.
Link:http://www.jbc.org/content/286/24/21588

2.Title:Bidirectional Concentration-Dependent Effects of Tumor Necrosis Factor Alpha in Shigella dysenteriae-Related Seizures.
Publication:INFECTION AND IMMUNITY, Apr. 2003, p. 2288–2291 Vol. 71, No. 40019-9567/03/$08.000 DOI: 10.1128/IAI.71.4.2288–2291.2003Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Link:http://iai.asm.org/content/71/4/2288.full.pdf

3.Title:Common Inhibitory Serine Sites Phosphorylated by IRS-1 Kinases, Triggered by Insulin and Inducers of Insulin Resistance.
Publication:First Published on April 19, 2007, doi: 10.1074/jbc.M610949200 June 22, 2007 The Journal of Biological Chemistry, 282, 18018-18027.
Link:http://www.jbc.org/content/282/25/18018.full

4.Title:Antioxidative function of L-FABP in L-FABP stably transfected Chang liver cells.
Publication:Article first published online: 20 SEP 2005 DOI:10.1002/hep.20857 Copyright © 2005 American Association for the Study of Liver Diseases.
Link:http://onlinelibrary.wiley.com/doi/10.1002/hep.20857/full

5.Title:Interferon-? induces apoptosis in human SH-SY5Y neuroblastoma cells through activation of JAK–STAT signaling and down-regulation of PI3K/Akt pathway
Publication:Article first published online: 11 NOV 2010 DOI:10.1111/j.1471-4159.2010.07046.x © 2010 The Authors. Journal of Neurochemistry © 2010 International Society for Neurochemistry.
Link:http://onlinelibrary.wiley.com/doi/10.1111/j.1471-4159.2010.07046.x/full

6.Title:Common Inhibitory Serine Sites Phosphorylated by IRS-1 Kinases, Triggered by Insulin and Inducers of Insulin Resistance.
Link:http://journal.9med.net/qikan/article.php?id=408762

7.Title:The human respiratory syncytial virus non-structural protein 1 regulates type I and type II interferon pathways.
Publication:First Published on February 8, 2012, doi: 10.1074/mcp.M111.015909 mcp.M111.015909.
Link:http://www.mcponline.org/content/early/2012/02/08/mcp.M111.015909.short

8.Title:Calcium transient evoked by TRPV1 activators is enhanced by tumor necrosis factor-? in rat pulmonary sensory neurons.
Publication:Published online before print July 16, 2010, doi: 10.?1152/?ajplung.?00111.?2010 AJP - Lung Physiol October 1, 2010 vol. 299 no. 4 L483-L492
Link:http://ajplung.physiology.org/content/299/4/L483.full

Usage Prospec's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.

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