Anti v5 agarose affinity gel

Western blot, immunoprecipitation and in vitro kinase assays were performed as previously described. The following antibodies were used: anti-p217/p221-MEK1/2 (p-MEK1/2)(#9154), anti-p202/p204-ERK1/2 (p-ERK1/2) (#4370), anti-MEK1/2 (#4694), anti-ERK1/2 (#4696) from Cell Signaling, anti-V5 (R960-25) from Thermo Fisher Scientific, anti-ARAF (sc-408) and anti-BRAF (sc-5284) from Santa Cruz Biotechnology, anti- FLAG (F3165) from Sigma, anti-CRAF (610152) from BD Transduction Laboratories, anti-p-CRAF S338 (05-538) from Millipore, and anti-RAS antibody from the active RAS pull-down and detection kit (Thermo Fisher Scientific, #16117). For immunoprecipitations of tagged proteins, the following reagents were used: anti-V5 agarose affinity gel (Invitrogen), anti-FLAG M2 affinity gel (Sigma), protein G agarose gel (Invitrogen).

Western blot, immunoprecipitation and in vitro kinase assays were performed as described (Poulikakos et al., 2011 (link)). The following antibodies were used: anti-p217/p221-MEK1/2 (p-MEK1/2), anti-p202/p204-ERK1/2 (p-ERK1/2), anti-MEK1/2, anti-ERK1/2 from Cell Signaling, anti-V5 from Invitrogen, anti-BRAF from Santa Cruz Biotechnology, anti-FLAG from Sigma, anti-CRAF from BD Transduction Laboratories, anti-CRAF-S338 from Millipore. For immunoprecipitations of tagged proteins, the following reagents were used: anti-V5 agarose affinity gel (Invitrogen), anti-FLAG M2 affinity gel (Sigma), protein G agarose gel (Invitrogene).