Binding of M-Ras to the Ras binding domains (RBD) of Raf-1 and Ral-GDS. Lysates of 293-cells transiently transfected with HA-tagged M-Ras wt, M-Ras G22V, N-Ras Q61K, or vector alone were mixed as indicated with GST-fusion proteins of the RBD of Raf-1 or Ral-GDS precoupled to glutathione beads. Proteins were eluted from the beads by boiling and, in parallel with samples of the original lysates (WCL), were subjected to SDS-PAGE and immunoblotting with anti-HA antibody 12CA5. (◂), HA-M-Ras; (◃), HA-N-Ras.