TNFa TNF alpha

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TNFa TNF alpha

 

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Tumor Necrosis Factor-Alpha (TNF-a), also known as cachectin, is named after its activity to cause tumor necrosis in vivo when injected into tumor-bearing mice. TNF-a is expressed as a 26 kDa membrane bound protein and is then cleaved by TNF-a converting enzyme (TACE) to release the soluble 17 kDa monomer which forms homotrimers in circulation. Recombinant TNF-a exists as homo-dimer, -trimer or -pentamer. TNF-a is believed to play roles in antitumor activity, immune modulation, inflammation, anorexia, cachexia, septic shock, viral replication and ematopoiesis. TNF-a is expressed in many types of cells but primarily in macrophage cells in response to immunological challenges such as bacteria (lipopolysaccharides), viruses, parasites, mitogens and other cytokines. TNF-a is closely related to the 25 kDa protein Tumor Necrosis Factor-b (lymphotoxin) with 28% amino acid sequence identity, sharing the same receptors (TNFR1 and TNFR2) and cellular actions. TNF-a causes cytolysis or cytostasis of many transformed cells, being synergistic with g -interferon in its cytotoxicity. Although it has little effect on many cultured normal human cells, TNF-a appears to be directly toxic to vascular endothelial cells. Other actions of TNF-a include stimulating growth of human fibroblasts and other cell lines, activating polymorphonuclear neutrophils and osteoclasts, and induction of interleukin-1, prostaglandin E2 and collagenase production. Although TNF-a is currently being evaluated in treatment of certain cancers and AIDS-related symptoms, the recombinant TNF-a offered by Biomyx is for research only.

 

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Recombinant Human Tumor Necrosis Factor a

 

CAS Number: 94948591

MDL Number: MFCD00148692

 

Catalog# Package size Price

T1100-20 20g $299

T1100-200 200g $750

T1100-1000 1.0mg $1999

*offer valid on second item of equal size when you purchase one item (catalog numbers T1100-20, -200 or -1000) at list price.

Formulation: Powder lyophilized from volatile buffer 50mM NH4HCO3 (pH 8.0).

Preservative: None.

MW: 17kDa

Purity: >97% (15% SDS-PAGE)

Source: Recombinant protein expressed in E. coli.

Sterility: 0.2 m membrane-filtered and packaged aseptically.

ED50*: 0.01-0.1ng/ml.

Endotoxin**: 0.1 EU/mg TNF-a

QC Tests: SDS-PAGE, Native PAGE, ELISA, Cytolysis, TC

 

 

Reconstitution and Use:

Reconstitute the contents of the vial using sterile phosphate-buffered saline (PBS) to a concentration no less than 100 mg/ml and aliquot for future use. (If the initial rehydration is too diluting, activity may be lost due to the non-specific adsorption to the container). The solution can then be further diluted to a working stock solution. Bovine serum albumin can be added to the working solution to protect TNF-a from loss at low concentrations.

 

If the product is going to be used for applications requiring absolute asepsis (e.g. cell culture), its best to filter-sterilize the solution using a sterile and non-pyrogenic 0.2m membrane before use.

 

Storage and Stability:

Upon receiving, store the product at -20 C. After reconstitution, store the working aliquots at 2-8 C for no more than 3 months. For extended storage, aliquot the rehydrated solution (100 mg/ml) and freeze at -70 C or -20 C. Avoid repeated freezing and thawing. More dilute solutions stored at -20 C will lose activity faster.

*Cytolysis Assay:

The ED50 is defined as the effective concentration of TNF-a that causes 50% cytolysis of murine L929B cells, a TNF-a sensitive mouse fibrosarcoma cell line, in the presence of actinomycin-D. Results may vary depending on cell line used. (Havell, E.A., 1987).

 

 

**Endotoxin Assay:

Endotoxin Unit (EU) is determined by Limulus Amebocyte Lysate (LAL) assay (Sigma).

 

Further information:

 

1)       Aggarwal, B., and Reddy, S., Nicola, N., ed., Tumor necrosis factor (TNF) Guidebook to Cytokines and Their Receptors , New York (1994), 103-104

2)       Beutler, B., Sporn, M., and Roberts, A, ed., cachectin/tumor necrosis factor and lymphotoxin Peptide Growth Factors and their Receptors II , New York (1991), 39-70

3)       Callard, R., and Gearing, A., The Cytokine Facts Book , New York (1994),

4)       Ruff, M.R. and G.E. Gifford, Lymphokines 2: 235, 1981.

5)       Eskandari, M.K., et al., Immunol. Invest. 19: 69-79, 1990.

6)       Havell, E.A., J. Immunol. 139: 4225-4231, 1987.

7)       Ware, C., et al., Thomson, A.W., ed., Tumor necrosis factor-related ligands and receptors. The Cytokine Handbook 3rd ed., , San Diego, CA (1998), 549

8)       Matthews, N., et al., Lymphokines and Interferons, A Practical Approach, Clemens, M., et al., eds. IRL Press, 221 (1987).

 

 

 

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