Mouse TIMP1/Metalloproteinase inhibitor 1 ELISA Kit

A sandwich ELISA from Assay Genie for the quantitative detection of mouse TIMP1 (Metalloproteinase Inhibitor 1). The assay uses a colorimetric, double-antibody format and is validated for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. TIMP1 is described as a key regulator of metalloproteinases involved in extracellular matrix remodeling, with imbalances linked to pathologies including cancer, inflammatory diseases, and fibrosis.

Performance characteristics include a sensitivity of 9.375 pg/mL and a detection range of 15.6-1000 pg/mL. Linearity was assessed in serum, EDTA plasma, and heparin plasma (n = 5) at dilution factors of 1:2, 1:4, and 1:8, with values of 87-104%, 83-101%, and 81-96% for serum; 89-98%, 82-101%, and 80-97% for EDTA plasma; and 87-105%, 85-98%, and 80-100% for heparin plasma. Recovery averaged 96% for serum (range 94-100%), 91% for EDTA plasma (range 86-104%), and 95% for heparin plasma (range 88-104%). The kit is stored at 2-8°C for 12 months.

The workflow involves equilibrating reagents and TMB substrate to room temperature, loading standards, samples, blanks, and a zero control onto the pre-coated plate, and incubating at 37°C for 90 minutes for antigen binding. A biotin-labelled detection antibody is then added and incubated at 37°C for 60 minutes, followed by HRP-Streptavidin (SABC) for 30 minutes at 37°C, TMB substrate development in the dark for 10-20 minutes, stop solution, and immediate absorbance reading at 450 nm. Sample preparation guidance covers serum (clot, centrifuge at 1000 × g for 20 minutes), plasma (anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2-8°C), tissue homogenate (homogenize in PBS with protease inhibitors), cell culture supernatant (centrifuge at 2500 rpm for 5 minutes), and cell lysate (lyse with protease inhibitors).