Western Blot: Use HRP Goat-anti-Rabbit IgG [H&L] (code R1364HRP) and TMB as asubstrate. A working dilution range of 1: 500 to 1: 2,000 is suggested for this application to detect TNFαfrom supernatants or lysates of 2 x 10^6 endotoxin-stimulated mouse peripheral bloodmononuclear cells (PBMC). PBMC are stimulated for 24 hours with 1 % (v/v) mouse serumplus 10 ng/mL E. coli LPS. Immunohistochemistry: A dilution range of 1: 400 is suggested for this immunoassay. Either paraffin fixation or cryofixation can be used for immunohistochemistry using adilution of 1: 200 for staining of TNFα.
Restrictions
For Research Use only
Concentration
1.1 mg/mL (by UV absorbance at 280 nm)
Buffer
0.02 M Potassium Phosphate, 0.12 M Sodium Chloride, pH 7.2, 0.01 % (w/v) Sodium Azide, None
Preservative
Sodium azide
Precaution of Use
This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store undiluted at 2-8 °C.
Liu, Zhang, Chen, Wang, Qiu, Liu, Wan, Guo: "Effects and mechanisms of alveolar type II epithelial cell apoptosis in severe pancreatitis-induced acute lung injury." in: Experimental and therapeutic medicine, Vol. 7, Issue 3, pp. 565-572, (2014) (PubMed).
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