ER alpha Antibody (sample size)

Figure 1. ChIP using the Diagenode monoclonal antibody against hERα
from ChIP assays were performed using MCF7 cells, treated with the ER agonist estradiol for 3 hours prior to harvesting., the Diagenode monoclonal antibody directed against ER alpha (Cat. No. MAb-009-050) and optimized primer sets for qPCR. Sheared chromatin from 3 million cells and 5 µg of antibody were used per ChIP experiment. Recovery (%: ChIP/input) and occupancy (x fold: +ve/-ve) are shown in figure 1. QPCR was performed with primers for the GREB1 promoter and for exon 2 of the myoglobin gene (Cat. No. pp-1006-050), used as a negative control. Figure 1 shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA) and the occupancy (ratio +/- control target). These results demonstrate the occupancy of the GREB1 promoter by ERalpha.

Figure 2. Western blot analysis using the Diagenode monoclonal antibody against hERα
Western blot analysis was performed on 100 fmol ER alpha (ERa) and ER beta (ERb1) recombinant protein with the Diagenode monoclonal antibody directed against ER alpha (Cat. No. MAb-009-050) at a concentration of 7 µg/ml. Figure 2 shows the specificity of the antibody for the ER alpha isoform, whereas the ER beta isoform is not recognized.

Figure 3. Immunocytochemistry using the Diagenode monoclonal antibody against hERα
COS-7 cells transiently overexpressing human ERα (left) or ERß1 (right) were labeled with the Diagenode antibody against ER alpha (Cat. No. MAb-009-050), used at a concentration of 15 µg/ml, followed by a biotinylated secondary antibody and peroxidase-labeled avidin. Figure 3 shows the specificity of the antibody for the ER alpha isoform.