Anti-Estrogen Receptor alpha antibody [SP1] (ab16660)

ab16660 staining Estrogen Receptor alpha in MCF7 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab16660 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

IHC image of ab16660 staining Estrogen Receptor alpha in normal human cervix formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16660, 1/250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human breast tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.