Human MFAP5 (Microfibrillar-associated protein 5) ELISA Kit

Sandwich ELISA kit from Assay Genie designed for the quantitative measurement of human Microfibrillar-associated protein 5 (MFAP5) in serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples. The assay uses a double-antibody sandwich format with colorimetric detection, a sensitivity of 0.188 ng/mL, and a working range of 0.313–20 ng/mL. MFAP5 is a component of elastin-associated microfibrils involved in the regulation of microfibril assembly and extracellular matrix organization, and it has been implicated in fibrosis, cancer, and cardiovascular disorders, making it a useful biomarker for research into these conditions.

Linearity for serum (n = 5) was 86–105% at 1:2, 86–101% at 1:4, and 88–98% at 1:8; EDTA plasma (n = 5) gave 84–98%, 85–98%, and 90–100% at the same dilutions; and heparin plasma (n = 5) gave 84–93%, 81–99%, and 84–96%. Recovery averaged 90% in serum (range 86–95%), 95% in EDTA plasma (85–103%), and 98% in heparin plasma (89–104%). Unopened kits are stored at 2–8°C for 12 months.

Sample preparation guidance includes allowing serum to clot and centrifuging at 1000 × g for 20 minutes, and collecting plasma in anticoagulant tubes and centrifuging at 1000 × g for 15 minutes at 2–8°C. Cell culture supernatant should be centrifuged at 2500 rpm for 5 minutes, while cell lysates are prepared using lysis buffer with protease inhibitors. The workflow is a standard sandwich ELISA: equilibrate reagents and TMB substrate to room temperature, load standards, samples and blanks and incubate at 37°C for 90 minutes, add biotinylated detection antibody and incubate for 60 minutes, add HRP-streptavidin (SABC) for 30 minutes, develop with TMB substrate for 10–20 minutes in the dark, stop the reaction, and read absorbance at 450 nm.