Antibody Evaluation for Protein Analysis

The monoclonal antibodies specific for α-Tubulin, E-Cadherin, AKT1, PARP1, HSC70, GADPH and acetylated-lysine and the polyclonal antibodies specific for ERβ, SIRT1, phospho-AKT (pSer473), AKT, were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-AKT1 and AKT3 monoclonal antibodies were from Rockland Immunochemicals Inc. (Gilbertsville, PA, USA). Anti-mouse and anti-rabbit IgG peroxidase conjugated antibodies and chemical reagents were from Sigma-Aldrich (St Louis, MO, USA). ECL, nitrocellulose membranes and protein assay kit were from Bio-Rad (Hercules, CA, USA). Culture media, sera, antibiotics and LipofectAMINE transfection reagent were from Invitrogen (Carlsbad, CA, USA). The AKT-inhibitor MK-2206 was obtained from Selleck Chemicals (Houston, TX, USA). The, previously described ERβ selective agonist KB9520 [30 (link), 45 (link)] was designed and synthesized by Karo Bio AB (Huddinge, Sweden).