Tumor Necrosis Factor alpha (TNFa)

Tumor Necrosis Factor alpha (TNF-alpha) is a 17.3kD multi-functional pro-inflammatory cytokine primarily secreted by macrophages, but also produced by T and B lymphocytes, activated monocytes and fibroblasts. TNF-alpha exists in both a transmembrane and mature soluble form and has many biological properties, including activation of transcription factor NF-KappaB, the induction of apoptosis in various tumor cell lines and the stimulation of cell differentiation and proliferation under certain conditions. Studies have revealed a role for TNF-alpha in inflammation, septic shock, autoimmune disease and rheumatoid arthritis. \n\nHybridoma:\nHybridization of P3X63-Ag8.653 myeloma cells with spleen cells from Balb/c mice.\n\nApplications: \nSuitable for use in Neutralization. Other applications not tested.\n\nRecommended Dilution:\nNeutralization: 250ng/ml of T9160-32 will neutralize 50% of the cytotoxic effect of 10ng/ml recombinant human TNFa, with an ED50 of 0.3ng/ml, on actinomycin D treated L929 cells. Previously, 500ng/ml of this antibody neutralized 50% of the cytotoxic effect of 10ng/ml recombinant human TNFa, with an ED50 of 0.04ng/ml. \nOptimal dilutions to be determined by the researcher.\n\nTNFa Neutralization Assay:\n1. Plate 2x10e4 murine L929B cells into individual wells of 96-well (flat bottomed) microtiter plate. \n2. Incubate overnight at 37 degrees C in humidified 5% CO2 incubator. \n3. The next day, prepare serial dilutions of T9160-32 in medium containing 2ug/ml of actinomycin D, for a final volume of 50ul. \n4. Add 50ul of TNFa (10ng/ml final concentration) to each antibody dilution and incubate for 1 hour at 37 degrees C. \n5. Add 100ul of the antibody/antigen mixture to the pre-seeded L929B cells. \n6. Incubate 24 hours at 37 degrees C, 5% CO2.\n7. Score wells for cytotoxicity.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol (40-50%), aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. 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.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts