Anti erk sc 93

Western blot analysis was done as previously described [41 (link)] by using the following antibodies: anti-hGH (National Hormone and Peptide Program, CA); anti-E-cadherin (ab1416, Abcam); anti-Fibronectin 1 (ab2413, Abcam); anti-Occludin (ab31721, Abcam); anti-p-ERK (sc-7383, Santa Cruz); anti-ERK (sc-93, Santa Cruz), and anti-β-actin (sc-58222, Santa Cruz). Immunofluorescence analysis was performed as previously described [89 (link)], and visualized under Nikon A1R-A1 Confocal Microscope System.

For SDS–PAGE analysis, the membranes were blocked with 5 % milk in PBS Tween 20 (10 %) for 30 min at room temperature. Membranes were then probed overnight at 4 °C with the appropriated primary antibodies: anti-MITF (HPA003259, Sigma), anti-HA (3F10, Roche), anti-myc (9E10, Santa Cruz), anti-flag (M2, Sigma), anti-ARAF (4432, Cell Signaling), anti-BRAF (sc5284, Santa Cruz), anti-CRAF (610151, BD Biosciences), anti-ERK (sc93, Santa Cruz), anti-pERK (M8159, Sigma), anti-laminA/C (10298-1-AP, Proteintech), anti-MEK1 (sc219, Santa Cruz) and anti-β-actin (A1978, Sigma) antibodies. Antigen-antibody complexes were detected by horseradish peroxidase-coupled secondary antibodies followed by enhanced chemiluminescence. Signals were acquired using a cooled-CDD camera (Fusion FX Spectra, Vilber).