For flow cytometry and Western blotting dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:10. For neutralization of biological activity in vitro dilutions have to be made according to the amounts of TNF-alpha to be inactivated. Before use in biological assays, the product must be filter sterilized and depending on the concentration to be used dialyzed against culture medium to remove the sodium azide added. Please inquire for availability of azide free solutions.
The antibody reacts with free soluble (17 kDa) and membrane (26 kDa) human TNF-alpha. The antibody inhibits the biological activity of both forms. It does not react with receptor bound TNF-alpha. It can be a useful tool to discriminate between the membrane form of TNF expressed on producer cells and the proteolytically cleaved, soluble TNF-alpha bound to its cognate cell membrane receptors (TNF-RI and TNF-RII). For this purpose we recommend to use this antibody in combination with the anti-TNF-alpha antibody HM2026, which recognizes soluble, membrane and receptor bound TNF-alpha.