Irdye 800cw anti rabbit and anti mouse secondary antibodies

Rabbit polyclonal antibody for phospho-p38MAPK [Thr 180/Tyr 182] and total-p38MAPK and mouse monoclonal antibody for CD36 were from Santa Cruz Biotechnology [Santa Cruz, CA]. Phospho-p53, total-p53 and cleaved caspase-3 antibodies were from Cell Signaling [Danvers, MA]. IRDye^® 800CW anti-rabbit and anti-mouse secondary antibodies were from LICOR [Lincoln, NE]. Metformin hydrochloride was purchased from Sigma-Aldrich. Rac1 antiserum was from BD Transduction Laboratories [San Jose, CA]. Antisera against cleaved caspase-3, lamin B, Phospho and total p53 and Myc tag were from Cell Signaling [Danvers, MA]. Mouse monoclonal β-actin antibody was obtained from Sigma-Aldrich [St. Louis, MO]. GGTI-2147 was from VDM biochemicals [Bedford Heights, OH]. EHT 1864 was from R&D Systems [Minneapolis, MN]. NE-PER^® Nuclear and cytoplasmic extraction kit was from Thermo Scientific [Waltham, CA]. Lipofectamine 3000 was from Invitrogen [Carlsbad, CA]. All other reagents used in the studies were obtained from Sigma-Aldrich [St. Louis, MO].

Cell proteomes were separated by SDS-PAGE, transferred to nitrocellulose membrane (60 V for 90 min), and blocked by 5% milk in TBS-Tween. The primary antibodies used and dilutions are as follows: anti-AIG1 (Rabbit, Sigma-Aldrich, SAB1304597, 1:250), anti-ADTRP (Rabbit, Atlas, HPA048113, 1:500), and anti-β-actin (Mouse, Cell Signaling, 3700S, 1:1000). IRDye 800CW anti-rabbit and anti-mouse secondary antibodies (LI-COR, 1:10000) were used as secondary antibodies for visualization.