Antibody-mediated Protein Analysis Protocol

The monoclonal antibodies specific for α-Tubulin, AKT1 and Acetylated-Lysine and the polyclonal antibodies specific for ERβ, phospho-AKT (pSer473), AKT, MCT1 and MCT4 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, 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 ERβ selective agonist KB9520 was designed and synthesized by Karo Bio (Huddinge, Sweden). (KB9520 has been described previously [32 (link), 45 (link), 46 (link)]. The compound can be obtained following contact with Karo Bio AB [stefan.nilsson@karobio.se] and after signing of a Material Transfer Agreement together with a detailed protocol of planned study. A fee covering the cost of compound synthesis will be charged).