Ab157712

CRP concentration in serum was measured using a quantitative sandwich ELISA (Abcam, ab157712) according to the manufacturer's protocol. Briefly, 100 μl of standards, control and serum samples (diluted 1:10 (v/v) in supplied diluent) from adult MSUS and control males were added to wells containing anti‐CRP. Then, HRP‐conjugated anti‐CRP antibodies were added. After 10 min of incubation at room temperature, wells were washed and then exposed to the chromogenic substrate tetramethylbenzidine (TMB) for 5 min in the dark at room temperature; then, a stop solution was added to each well. Absorbance was measured at 450 nm with a NOVOstar Plate Reader (BMG Labtech), and sample absorbance (directly related to concentration) was determined by plotting a standard curve with manufacturer‐supplied standards and controls. Concentrations were analysed with one‐tailed Student's t‐test for mouse samples (as validation of proteomic data).