Rabbit polyclonal anti human p erk

For Western blot analysis, cells or exosomal preparations were lysed using lysis buffer (50 mmol/L Tris-HCl, pH 7.2, 150 mmol/L NaCl, 100 mmol/L NaF, 100 mmol/L sodium pyruvate, and 1% Triton X-100) containing protease inhibitors, 2 mmol/L phenyl methyl sulfonyl fluoride, 10 mg/ mL aprotinin, and 2 mmol/L Na 3 VO 4 . Extracted protein (10 mg) was separated by SDS-PAGE, transferred to immobilon-P membranes, and analyzed using the enhanced chemiluminescence kit for Western blotting detection (Amersham Pharmacia Biotech, Milan, Italy), as previously described. 15 Primary monoclonal antibodies were used following suppliers' instructions and included the following: mouse anti-human monoclonal CD63 (dilution, 1:1000; Santa Cruz Biotechnology, Inc., Dallas, TX), mouse monoclonal anti-human CD133 (dilution, 1:200; Miltenyi Biotec S.r.l., Bologna, Italy), rabbit polyclonal anti-human caveolin-1 (Cav-1; dilution, 1.500; Santa Cruz Biotechnology, Inc.), rabbit polyclonal anti-human Src (dilution, 1:1000; Cell Signaling Technology, Milan, Italy), rabbit polyclonal antihuman pSrc (dilution, 1:1000; Cell Signaling Technology), rabbit polyclonal anti-human extracellular signal regulated kinase (ERK; dilution, 1:1000; Cell Signaling Technology), and rabbit polyclonal anti-human pERK (dilution, 1:1000; Cell Signaling Technology).