Western blot using Gdf15 polyclonal antibody (Cat # PAB16912). Lane 1, recombinant MBP-Gdf15 fusion protein (60 KDa) purified from E.coli. Lane 2, yeast cell lysate expressing SUMO-mouse Gdf15 (42 KDa). Lane 3, R&D human Gdf15 monomer purified from CHO-K1 cells (14 KDa). All lysates were run under reducing conditions. Primary antibody was used at a 1 : 1000 dilution in TBS containg 1% BSA and 0.2% Tween, and reacted overnight at 4°C. Gdf15 was detected using a 1 : 40,000 dilution of peroxidase conjugated Gt-a-Rabbit antibody in Blocking Buffer for Fluorescent Western Blotting for 30 min at room temperature. Molecular weight estimation was made by comparison to prestained MW markers. Image was captured using the BioRad Versadoc™4000MP Imaging System.