85-7206-12　Tumor Necrosis Factor alpha (TNFa) 1mg　T9160-31

特徴

• 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.
• Applications:Suitable for use in Dot Blot and Neutralization.
• Other applications have not been tested.
• Recommended Dilutions:Dot Blot: 1ug/ml detected 5ng recombinant murine TNFa.
• Neutralization: 3ug/ml neutralized 50% of the cytoxic effect of 50pg/ml recombinant mouse TNFa on actinomycin D treated L929 cells.
• The antibody concentation needed to neutralize the bioactivity of TNFa will depend on cell type, incubation conditions and biological response.
• Optimal dilutions to be determined by the researcher.
• Storage and Stability:May be stored at 4°C for short-term only.
• Aliquot to avoid repeated freezing and thawing.
• Store at -20°C.
• Aliquots are stable for 12 months after receipt.
• For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
• TNF-a Neutralization Assay:1.
• Plate 2 x 10e4 murine L929B cells into individual wells of 96-well (flat bottomed) microtiter plate.
• 2.
• Incubate overnight at 37°C in humidified 5% CO2 incubator.
• 3.
• The next day, prepare serial dilutions of T9160-31 in medium containing 2ug/ml of actinomycin D, for a final volume of 50ul.
• 4.
• Add 50ul of mouse TNFa (50pg/ml final concentration) to each antibody dilution and incubate for 1 hour at 37°C.
• 5.
• Add 100ul of the antibody/antigen mixture to the pre-seeded L929B cells.
• 6.
• Incubate 24 hours at 37°C, 5% CO2.
• 7.
• Score wells for cytotoxicity.