ELISA Capture: Purified G281-2626 antibody is useful as a capture antibody for a sandwich ELISA for measuring either mouse or rat TNF protein levels. The purified G281-2626 antibody should be titrated from 4-8 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of TNF protein (Cat. No. 554589) ranging from 4,000 to 20 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, visit the protocols section of our website, or the chapter on ELISA in the Immune Function Handbook, which is posted on our web site at www.bdbiosciences.com.
MOUSE TNF ELISA: The combination of purified G281-2626 antibody with biotin MP6-XT3 (Cat. No. 554415) is useful for measuring free TNF. The combination of purified TN3-19.12 (Cat No. 557516) with biotin C1150-14 (Cat No. 557432) is useful for measuring total TNF, both free TNF as well as TNF bound by soluble TNF receptors.
Note: These ELISA pairs are recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring free TNF in serum or plasma our anti-mouse TNF BD OptEIA™ ELISA Set (Cat. No. 555268) is specially formulated and recommended. For measuring total TNF in serum or plasma our anti-mouse TNF OptEIA Set (Cat. No. 558534) or OptEIA Kit (Cat. No. 559732) is specially formulated and recommended.
RAT TNF ELISA: Purified G281-2626 antibody can be paired with the biotinylated IgG fraction of polyclonal rabbit anti-mouse/rat TNF antiserum (Cat. No. 557432) as the detecting antibody and with recombinant rat TNF (Cat. No. 555109) as the standard.
Note: These ELISA pairs are recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring rat TNF in serum or plasma, a our anti-rat TNF OptEIA Set (Cat. No. 558535) is specially formulated and recommended.
Please note: For best results, use 0.1 M sodium phosphate pH 6.0 binding buffer with this antibody.
Western blotting: The G281-2626 antibody has been found useful for Western blotting; however, the antibody is not tested for this application at BD Biosciences PharMingen.