Immunoprecipitation: mTOR Antibody - Several commercially available antibodies which were generated using short peptides failed to isolate the FRAP1 protein complexes. SDI's FRAP1 (mTOR) antibody, 2813.00.02, was generated using in vivo expressed immunogen of 100 amino acids. It was used to pull down protein complexes through immunoprecipitation. It demonstrated that Rictor and Raptor formed the distinct complexes with FRAP (mTOR). The immunoprecipitation conditions were described in Sarbassov et al (Phosphorylation and regulation of Akt/PKB by the Rictor-mTOR complex, Science 307:1098-1101, 2005, supporting online material, material and methods). FRAP1 was detected in Western blot using FRAP1 antibody at 1:4000 dilution. The experiments were performed using MDA MB-435 human cancer cell line and NF2 was used as control.