Quantification of Soluble Mertk in Atherosclerosis

Mertk ELISA was conducted on supernatants of BMDMs or sections of atherosclerotic plaques lysed in protein extraction buffer (300 mM Tris-HCl pH 8.0 and 2% SDS) (Kawashima et al., 2014 (link)). Anti-Mertk antibody (AF591, R&D Systems) was used as the capture antibody. 200 µL of culture supernatant or 10 µg equivalent of protein from plaque lysates were loaded per well. Biotinylated anti-Mertk antibody (BAF591, R&D Systems) was used as a detection antibody followed by incubation with streptavidin-HRP and colorimetric development using TMB substrate. A450_nm was used to estimate the quantity of soluble Mertk.