Flag-EMB140 expressed in HEK293T cells was added to a kernel extract similar to that used in the in vitro stability assay for ZmEMB140 proteins, supplemented with a final concentration of 50mM Mg132. After incubation at room temperature for 1 hour, Flag-EMB140 was precipitated using Flag beads. Following this, SDS-loading buffer was added and the proteins were separated via SDS-PAGE gel electrophoresis