Description
The G281-2626 antibody reacts with mouse and rat tumor necrosis factor (TNF). The immunogen used to generate the G281-2626 hybridoma was recombinant mouse TNF. This is a neutralizing antibody. Cross reactivity with LT-α has not been determined.
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.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
