This MAb recognizes human 17-26kDa protein, which is identified as cytokine TNF-alpha (Tumor Necrosis Factor-alpha). TNF-alpha can be expressed as a 17kDa free molecule, or as a 26kDa membrane protein. TNF-alpha is a protein secreted by lipopolysaccharide-stimulated macrophages, and causes tumor necrosis when injected into tumor 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 non-covalently linked units, but shows a single 17kDa band following SDS PAGE under non-reducing conditions. TNF alpha is closely related to the 25kDa protein Tumor Necrosis Factor beta (lymphotoxin), sharing the same receptors and cellular actions. TNF alpha causes cytolysis 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.
Suitable for use in Immunofluorescence, ELISA, Western Blot, Immunoprecipitation, Immunohistochemistry. Other applications not tested.
Immunoprecipitation: 1-2ug/500ug protein
Western Blot: 1-2ug/ml
Immunohistochemistry: Frozen: 1-2ug/ml for 30 min at RT
Optimal dilutions to be determined by the researcher.
HeLa, HL-60, or A431 cells. Macrophages in lymph node or Tonsil
Storage and Stability:
May be stored at 4°C before opening. DO NOT FREEZE! Stable at 4°C as an undiluted liquid. Dilute only prior to immediate use. Stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Freezing alkaline phosphatase conjugates will result in a substantial loss of activity.
Note: Applications are based on unconjugated antibody.