Human Thyroid peroxidase/TPO ELISA Kit

From Assay Genie, this Sandwich ELISA kit uses a double antibody format for the quantitative measurement of Thyroid peroxidase/TPO (also referred to by the synonyms TPO, Thrombopoietin, MGDF, MK-CSF and THPO) in human samples. The assay is intended for signal transduction research and accepts serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. Detection is colorimetric, with a sensitivity of 18.75 pg/mL and a working range of 31.25–2000 pg/mL.

Analytical performance has been characterised across serum, EDTA plasma and heparin plasma. Linearity at 1:2, 1:4 and 1:8 dilutions returned values of approximately 85–103%, 86–99% and 86–104% in serum, 84–101%, 93–98% and 87–100% in EDTA plasma, and 85–96%, 82–99% and 80–97% in heparin plasma. Spike-and-recovery across the same three matrices ranged from 85–99% (average 92%) in serum, 86–101% (average 94%) in EDTA plasma and 87–103% (average 94%) in heparin plasma. Unopened components are stored at 2–8°C for 12 months.

The assay workflow involves equilibrating reagents and TMB substrate to room temperature; a primary incubation at 37°C for 90 minutes to allow antigen binding; incubation with a biotin-labelled detection antibody at 37°C for 60 minutes; incubation with HRP-Streptavidin (SABC) at 37°C for 30 minutes; colour development with TMB substrate in the dark for 10–20 minutes; and a stop reaction with absorbance measured immediately at 450 nm. Sample preparation differs by matrix: serum is allowed to clot and centrifuged at 1000 × g for 20 minutes; plasma is collected into anticoagulant tubes and centrifuged at 1000 × g for 15 minutes at 2–8°C; tissue is homogenised in PBS with protease inhibitors; cell culture supernatant is clarified at 2500 rpm for 5 minutes; and cell lysates are prepared with a lysis buffer containing protease inhibitors.