Mouse TWEAK (Tumour Necrosis Factor Related Weak Inducer of Apoptosis) ELISA Kit

A colorimetric sandwich ELISA from Assay Genie designed for the quantitative detection of mouse TWEAK (Tumour Necrosis Factor Related Weak Inducer of Apoptosis). The assay uses a double-antibody sandwich format and is validated for use with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Performance specifications include a sensitivity of 9.375 pg/mL and a working range of 15.625–1000 pg/mL. Reagents are stored at 2–8°C for 12 months.

The standard protocol comprises six steps: reagent and plate preparation at room temperature, primary incubation at 37°C for 90 minutes, biotinylated detection antibody binding at 37°C for 60 minutes, HRP-Streptavidin (SABC) binding at 37°C for 30 minutes, TMB colour development in the dark for 10–20 minutes, and stop reaction with absorbance reading at 450 nm. Sample preparation guidance is provided for serum (centrifuge at 1000 × g for 20 minutes), plasma (centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate (homogenise in PBS with protease inhibitors), cell culture supernatant (centrifuge at 2500 rpm for 5 minutes) and cell lysate (lyse with lysis buffer containing protease inhibitors).

Reported linearity is serum (n=5) 86–105% at 1:2, 87–103% at 1:4, 85–104% at 1:8; EDTA plasma (n=5) 83–96% at 1:2, 84–101% at 1:4, 84–101% at 1:8; heparin plasma (n=5) 82–92% at 1:2, 81–90% at 1:4, 82–94% at 1:8. Reported recovery is serum (n=5) 93–102% range with 99% average, EDTA plasma (n=5) 88–99% range with 95% average, and heparin plasma (n=5) 89–101% range with 95% average. The associated Uniprot identifier is O54907.