Analyzing Gas6-Mer/Axl Signaling Pathways

Reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA) unless otherwise stated. DMEM and Iscove's DMEM (IDMEM) were from Invitrogen (Carlsbad, CA, USA). Human Protein S was obtained from Enzyme Research Laboratories (South Bend, IN, USA). Recombinant mouse Gas6 (full-length and Gla-less) was prepared as previously described.^{24 , 48 (link)} BMS777607 was obtained from Selleckchem (Houston, TX, USA). Goat polyclonal antibodies against human Mer and mouse Mer and Axl and were obtained from R&D Systems (Minneapolis, MN, USA). Monoclonal anti-human Mer (mouse IgG1) was from R&D Systems, anti-GAPDH (mouse IgG1) was from Millipore (Billerica, MA, USA), anti-mouse CD61 mAb (Hamster, Hmbeta 3.1) and TIM4 (Rat IgG2a – clone 54) were obtained from eBioscience (San Diego, CA, USA). Control immunoglobulins, and secondary HRP or fluorescent-conjugated antibodies were from DAKO (Carpinteria, CA, USA). All antibodies were titrated on cells or cell lines known to express the antigen. Enhanced chemiluminescence (ECL) substrate and Hyperfilm was obtained from GE Healthcare (Pittsburgh, PA, USA).