This IgG fraction of anti-Human TNFa has been tested for use in Neutralizations, ELISA(1/1,000-1/5,000), Immunohistochemistry, Radioimmunoassay, Immunoprecipitation,Immunohistochemistry and Immunoblotting (1/500-1/2,000). It recognizes the 17,000 MWTNFa . Reactivity in other immunoassays is unknown. Recommended Dilutions: Immunoblot using HRP Goat-anti-Rabbit IgG [H&L] and TMB as a substrate. A workingdilution range of 1/100-1/200 is suggested for this application to detect TNFa fromsupernatants or lysates of 2 x 10^6 endotoxin-stimulated human peripheral bloodmononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1 % (v/v) human serumplus 10 ng/mL E. coli LPS.
Restrictions
For Research Use only
Concentration
5.0 mg/mL (by UV Absorbance at 280 nm)
Buffer
0.02 M Potassium Phosphate, 0.12 M Sodium Chloride, pH 7.2 without stabilizers or preservatives.
Preservative
Without preservative
Handling Advice
Avoid repeated freezing and thawing. This product is photosensitive and should be protected from light.
Storage
4 °C/-20 °C
Storage Comment
Store the antibody undiluted at 2-8 °C for one month or (in aliquots) at -20 °C for longer.
Tumor Necrosis Factor Alpha (TNF alpha) is a protein secreted by lipopolysaccharide stimulated macrophages, and causes tumor necrosis when injected into tumour bearing mice. TNF alpha is believed to mediate pathogenic shock and tissue injury associated with endotoxemia. TNF alpha exists as a multimer of two, three, or five noncovalently linked units, but shows a single 17 kDa band following SDS PAGE under non reducing conditions. TNF alpha is closely related to the 25 kDa protein Tumour Necrosis Factor beta (lymphotoxin), sharing the same receptors and cellular actions. TNF alpha causes cytolysis or cytostasis of certain transformed cells, being synergistic with interferon gamma in its cytotoxicity. Although it has little effect on many cultured normal human cells, TNF alpha appears to be directly toxic to vascular endothelial cells. Other actions of TNF alpha 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. TNF alpha is currently being evaluated in treatment of certain cancers and AIDS Related Complex.Synonyms: Cachectin, TNF, TNF-a, TNFA, TNFSF2, Tumor necrosis factor ligand superfamily member 2