Bovine TNF-alpha (Tumor necrosis factor alpha) ELISA Kit

From Assay Genie, this sandwich double-antibody ELISA kit enables quantitative measurement of Bovine TNF-alpha (Tumor necrosis factor alpha). The assay uses a microplate pre-coated with a capture antibody specific to the target protein, with colourimetric detection via HRP-conjugated Streptavidin and TMB substrate. Standards and appropriately diluted samples are incubated in the wells; after washing, a biotinylated detection antibody binds a different epitope of the analyte to form the sandwich complex. The colour reaction is stopped with an acidic solution and absorbance is read at 450 nm, with signal intensity proportional to analyte concentration as determined from a standard curve.

The kit is reactive with Bovine samples and is validated for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Reported sensitivity is 9.375 pg/mL with a working range of 15.625–1000 pg/mL. Storage is 2–8°C for 12 months.

Linearity data (n=5) for serum spans 89–101%, 85–103%, and 85–104% at 1:2, 1:4, and 1:8 dilutions respectively. EDTA plasma shows 86–95%, 87–97%, and 85–101% across the same dilutions, while heparin plasma gives 87–100%, 83–100%, and 81–98%. Recovery is 89–104% (serum, mean 95%), 88–99% (EDTA plasma, mean 95%), and 90–105% (heparin plasma, mean 98%).

The protocol includes primary incubation at 37°C for 90 minutes, biotinylated detection antibody binding at 37°C for 60 minutes, HRP-Streptavidin incubation at 37°C for 30 minutes, and TMB colour development in the dark for 10–20 minutes.