Human Trypsin ELISA Kit

Designed for the quantitative measurement of human trypsin, this sandwich double-antibody ELISA from Assay Genie uses a colorimetric detection method with absorbance read at 450 nm. It is compatible with serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The microplate is pre-coated with a capture antibody specific to the target, and a biotinylated detection antibody directed to a different epitope forms the sandwich complex. HRP-conjugated streptavidin and TMB substrate generate the colorimetric signal, which is stopped with an acidic solution and quantified against a standard curve.

The reported sensitivity is 0.938 ng/mL with a measuring range of 1.563–100 ng/mL, and kits are intended to be stored at 2–8°C for 12 months. The protocol includes equilibrating reagents to room temperature; loading standards, samples and blanks onto the pre-coated plate; incubating at 37°C for 90 minutes; adding the biotin-labeled detection antibody with a 60-minute incubation at 37°C; adding HRP-Streptavidin (SABC) for 30 minutes at 37°C; developing with TMB substrate in the dark for 10–20 minutes; and reading absorbance at 450 nm immediately after adding the stop solution.

Reported linearity for serum (n=5) is 85–95% at 1:2, 83–99% at 1:4 and 86–93% at 1:8; for EDTA plasma (n=5), 85–98%, 85–96% and 94–100%; and for heparin plasma (n=5), 89–103%, 83–98% and 82–90%. Recovery averages are 98% for serum, 94% for EDTA plasma and 96% for heparin plasma. Sample preparation guidance includes clotting and centrifuging blood at 1000 × g for 20 minutes for serum, collecting plasma using anticoagulant tubes centrifuged at 1000 × g for 15 minutes at 2–8°C, homogenising tissue in PBS with protease inhibitors, centrifuging cell culture supernatant at 2500 rpm for 5 minutes, and lysing cells using lysis buffer with protease inhibitors.