Mouse axl duoset elisa

Mer and Axl were measured using Mouse Mer and Mouse Axl DuoSet ELISA (R&D Systems). To measure Protein S in plasma, polystyrene High Bind Microplates (Corning) were coated with a polyclonal anti-human Pros1 antibody (Dako) in 75 mM Na-carbonate pH 9.6 o/n at +4 °C. In between each assay step, plates were washed 3× with PBS containing 0.1% Tween 20. Wells were blocked with 2 % BSA in PBS, which was also used as sample and antibody dilution buffer. A standard curve was generated with a pooled plasma sample from 11 mice, and individual samples diluted at 1:400 were analyzed. Samples and standards were incubated on the plate for 2 h at RT, followed by incubation with a rat anti-mouse Pros1 antibody (R&D Systems) for 1 h. After an additional 1 h incubation with a goat anti-rat HRP conjugate (Jackson ImmunoResearch), the plates were developed with a TMB substrate (R&D Systems).

sAXL levels were determined in mouse serum samples by specific sandwich ELISA using commercial kits (DY854, mouse AXL DuoSet ELISA, R&D, Minneapolis, MN, USA) following the manufacturer’s instructions.