Human TWEAK R/TNFRSF12 ELISA Kit

From Assay Genie, this Sandwich ELISA kit enables quantitative detection of human TWEAK R/TNFRSF12 (UniProt Q9NP84), also referred to as TNFRSF12A, Fn14, CD266, CD266 antigen, FGF-inducible 14, fibroblast growth factor-inducible immediate-early response protein 14, and tumor necrosis factor receptor superfamily member 12A. The assay uses a colorimetric, double-antibody sandwich format and is intended for use with human samples including serum, plasma, cell culture supernatant, cell or tissue lysates, and other liquid samples. The product is positioned for cardiovascular research applications.

Linearity across 1:2, 1:4 and 1:8 dilutions spans 89–105% for serum, EDTA plasma and heparin plasma. Sample recovery ranges are 95–99% for serum (average 97%), 95–101% for EDTA plasma (average 98%), and 95–105% for heparin plasma (average 100%). Reagents are stored at 2–8°C for 12 months, with some components also suitable for 12 months at –20°C, and light-sensitive reagents protected from direct light.

The procedure comprises reagent and plate preparation, primary incubation at 37°C for 90 minutes, biotin-labelled detection antibody binding at 37°C for 60 minutes, HRP-Streptavidin binding at 37°C for 30 minutes, TMB colour development in the dark for 10–20 minutes, and absorbance reading at 450 nm after adding the stop solution. Sample preparation covers serum clotting and centrifugation at 1000 × g for 20 minutes, plasma collection in anticoagulant tubes with centrifugation at 1000 × g for 15 minutes at 2–8°C, tissue homogenisation in PBS with protease inhibitors, cell culture supernatant clarification at 2500 rpm for 5 minutes, and cell lysis with protease-inhibitor-containing buffer.