Bcakground

Tumor necrosis factor alpha (TNF-alpha ), also known as cachectin and TNFSF2, is the prototypic ligand of the TNF superfamily. It is a pleiotropic molecule that plays a central role in inflammation, immune system development, apoptosis, and lipid metabolism (1, 2). Mouse TNF-alpha consisits of a 35 amino acid (aa) cytoplasmic domain, a 21 aa transmembrane segment, and a 179 aa extracellular domain (ECD) (3). Within the ECD, mouse TNF-alpha shares 94% aa sequence identity with rat and 70%-77% with bovine, canine, cotton rat, equine, feline, human, porcine, rat, and rhesus TNF-alpha. TNF -alpha is produced by a wide variety of immune, epithelial, endothelial, and tumor cells (1, 2). TNF-alpha is assembled intracellularly to form a noncovalently linked homotrimer which is expressed on the cell surface (4). Cell surface TNF-alpha can induce the lysis of neighboring tumor cells and virus infected cells, and it can generate its own downstream cell signaling following ligation by soluble TNFR I (2, 5). Shedding of membrane bound TNF-alpha by TACE/ ADAM17 releases the bioactive cytokine, a 55 kDa soluble trimer of the TNF-alpha extracellular domain (6-8). TNF-alpha binds the ubiquitous 55-60 kDa TNF RI (9, 10) and the hematopoietic cell-restricted 80 kDa TNF RII (11, 12), both of which are also expressed as homotrimers (1, 2, 13).

Reference

1. Zelova, H. and J. Hosek (2013) Inflamm. Res. 62:641.

2. Juhasz, K. et al. (2013) Expert Rev. Clin. Immunol. 9:335.

3. Fransen, L. et al. (1985) Nucleic Acids Res. 13:4417.

4. Tang, P. et al. (1996) Biochemistry 35:8216.

5. Perez, C. et al. (1990) Cell 63:251.

6. Black, R.A. et al. (1997) Nature 385:729.

7. Moss, M.L. et al. (1997) Nature 385:733.

8. Gearing, A.J.H. et al. (1994) Nature 370:555.

9. Schall, T.J. et al. (1990) Cell 61:361.

10. Loetscher, H. et al. (1990) Cell 61:351.

11. Dembic, Z. et al. (1990) Cytokine 2:231.

12. Smith, C.A. et al. (1990) Science 248:1019.

13. Loetscher, H. et al. (1991) J. Biol. Chem. 266:18324

Specifications

Synonyms	APC1 protein; Cachectin; Cachetin; DIF; TNF; TNF, monocyte-derived; TNFA; TNF-A; TNFalpha
Accession #	P06804
Source	Human embryonic kidney cell, HEK293-derived mouse TNF-alpha protein
Source	Leu80-Leu235
Predicted Moleucular weight	17 kDa

Components and Storage

Formulation	Solution protein
	Dissolved in sterile PBS buffer.
	This solution can be diluted into other aqueous buffers. Centrifuge the vial prior to opening.
Storage and Stability	Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
	It is recommended that the protein be aliquoted for optimal storage
	12 months from date of receipt, -20 to -70 °C as supplied.
Shipping	Shipping with dry ice.

Quality

Purity	> 95%, determined by SDS-PAGE.
Endotoxin Level	<0.010 EU per 1 ug of the protein by the LAL method.
Activity	Measured in a cytotoxicity assay using L-929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. Matthews, N. and M.L. Neale (1987) in Lymphokines and Interferons, A Practical Approach.
Activity	The EC50 for this effect is 5-20 pg/mL.

SDS-PAGE

Gel filtration

Bioactivity

Documents

Mouse TNFa