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Synthetic peptide corresponding to Human Estrogen Receptor alpha aa 21-32.
Our Abpromise guarantee covers the use of ab3575 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 0.5 mg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 64 kDa (predicted molecular weight: 68 kDa).Can be blocked with Human Estrogen Receptor alpha peptide (ab5847).|
|Flow Cyt||Use at an assay dependent concentration. PubMed: 23768049Ab171870-Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.|
|ICC||Use at an assay dependent concentration.|
|IHC-P||Use a concentration of 1 - 5 µg/ml.|
ab3575 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab3575 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of Estrogen Receptor alpha staining in Human breast Ductal carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab3575, 5µg/ml, 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.
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.
ab3575 at a dilution of 1-5µg/ml staining ER alpha in Rat prostate tissue sections.
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