Localization of SMC4 to Mitotic Chromosomes by Immunofluorescence. Samples: HeLa cells grown on coverslips, fixed with methanol/acetone and blocked with 3% BSA in PBS. Antibody: Affinity purified rabbit anti-SMC4 antibody used at 5 ug/ml. Detection: Rhodamine labeled anti-rabbit secondary antibody. Cells were counterstained with DAPI.
Detection of Human SMC4 by Western Blot. Samples: Whole cell lysate (90 ug) from 293T cells (lane 1), SMC4 transfected 239T cells (lane 2) or normal human lymphoblasts (lane 3) separated on an 8% SDS-PAGE gel. Antibodies: Affinity purified rabbit anti-SMC4 used 0.2 ug/ml and goat anti-rabbit HRP used at a dilution of 1:20000. Detection: Chemiluminescence with 5 second.