ELISA: The purified TN3-19.12 antibody (Cat. No. 557516) can be used as a capture antibody for a sandwich ELISA that measures TNF protein levels:
MOUSE TNF ELISA: The purified TN3-19.12 capture antibody (Cat. No. 557516) can be paired with the biotinylated Fab anti-Mouse/Rat TNF (Cat. No. 558415) as the detection antibody and with recombinant mouse TNF protein (Cat. No. 554589) as the cytokine standard in a sandwich ELISA for measuring mouse TNF protein levels.
RAT TNF ELISA: The purified TN3-19.12 capture antibody (Cat. No. 557516) can be paired with the biotinylated Fab anti-Mouse/Rat TNF (Cat. No. 558415) and with recombinant rat TNF protein (Cat. No. 555109) as the cytokine standard in a sandwich ELISA for measuring rat TNF protein levels.
The capture and detecting antibody preparations should be titrated from 0.5 - 2 µg/ml to determine their optimal concentrations for ELISA. To obtain linear standard curves, doubling dilutions of TNF protein ranging from ~2,000 to 5 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. These ELISA antibody pairs show no cross-reactivity with other tested cytokines including mouse IL-1α, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3 and human: TNF.
Note: The BD OptEIA™ ELISA Kits and Sets are specially formulated for cytokine measurements in serum or plasma samples. For mouse TNF, the Mouse TNF OptEIA Set, Cat. No. 558534 and Mouse TNF OptEIA Kit, Cat. No. 559732 is recommended. For rat TNF, the Rat TNF OptEIA Set, Cat. No. 558535 and the Rat TNF OptEIA Kit 550734 are recommended.
In vitro neutralization: The NA/LE™ format of the TN3-19.12 antibody (Cat. No. 557370) is useful for neutralization of mouse TNF bioactivity.
Blocking Control for Intracellular Staining: The purified TN3-19.12 antibody can be used as a blocking control to demonstrate specificity of TNF staining by conjugated TN3-19.12 antibodies.
IP/WB: The TN3-19.12 antibody has been reported to be useful for immunoprecipitation and Western blotting. Please note that these applications are not routinely tested at BD Biosciences.