Western blot analysis of extracts from various cell lines and tissues using Estrogen Receptor β Antibody.
|REACTIVITY||H M R Mk|
|MW (kDa)||52, 55, 63|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody datasheet or product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised November 2013
Reprobing of an existing membrane is a convenient means to immunoblot for multiple proteins independently when only a limited amount of sample is available. It should be noted that for the best possible results a fresh blot is always recommended. Reprobing can be a valuable method but with each reprobing of a blot there is potential for increased background signal. Additionally, it is recommended that you verify the removal of the first antibody complex prior to reprobing so that signal attributed to binding of the new antibody is not leftover signal from the first immunoblotting experiment. This can be done by re-exposing the blot to ECL reagents and making sure there is no signal prior to adding the next primary antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
posted June 2005
revised October 2016
Protocol Id: 10
Estrogen Receptor β Antibody detects endogenous levels of total Estrogen Receptor β protein. This antibody is predicted to cross-react with all Estrogen Receptor β isoforms. This antibody does not cross-react with Estrogen Receptor α.Species Reactivity:
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Estrogen Receptor β1 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Estrogen Receptor β (ER β) is a member of the nuclear receptor superfamily of transcription factors and was discovered to be encoded by a gene (ESR2) distinct from that encoding Estrogen Receptor α (ER α) (1,2). While studies have revealed that alternative splicing generates mutiple isoforms of ER β that differ at their respective C-termini and in tissue distribution, ER β1 is believed to be the longest and only fully functional isoform (3,4). Indeed, it has been reported that shorter isoforms of ER β (ER β2, β4, and β5) can heterodimerize with ER β1 and enhance its transcriptional activity in an estradiol-dependent manner (4). ER β is expressed in a wide range of normal and malignant tissues, many of which coexpress ER α. It is proposed that ER β has an antiproliferative role, perhaps through heterodimerization with ER α and repression of its transcriptional activity at estrogen response elements (5,6). Recent studies have revealed that expression of ESR2 is subject to epigenetic regulation and that loss of ER β expression positively contributes to epithelial-mesenchymal transition and enhanced invasiveness in prostate cancer (7,8). ER β has also been found to be negatively regulated at the posttranslational level through phosphorylation of its AF-1 domain, which promotes its ubiquitin-dependent proteasomal degradation (9,10).
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