Brca1 d 9 sc 6954

The transfection experiments described under Transfection and cell cultivation were scaled to a 12-well plate setup for expression analysis of the GAL4 DBD:BRCA1(aa 1396–1863) fusion protein constructs by western blot analysis. Ten microgramme of sample protein was run on a 10% Mini-PROTEAN® TGX™ gel (Bio-Rad Laboratories, Hercules, CA, USA) and blotted onto 0.2 μm Nitrocellulose Membranes (Bio-Rad Laboratories, Hercules, CA, USA). The membranes were blocked with 5% BSA. Primary immunoblot staining was done using BRCA1 (D-9):sc-6954 (Santa Cruz Biotechnology, Dallas, TX, USA) in a 1:1000 dilution, overnight at 4 °C, and followed by staining with secondary antibody m-IgGκ BP-HRP: sc-516102 (Santa Cruz Biotechnology, Dallas, TX, USA) (1:1000) for 1 h at room temperature, before exposure using the ECL™ Prime Western Blotting Detection Reagent (GE Healthcare, Buckinghamshire, UK) with ImageQuant™ LAS 4000 and ImageQuant™ TL 1D v8.1 (GE Healthcare, Buckinghamshire, UK).