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49757
Estrogen Receptor α Activation Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

Estrogen Receptor α Activation Antibody Sampler Kit #49757

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Estrogen Receptor α Activation Antibody Sampler Kit: Image 1
Western blot analysis of extracts from untransfected MCF-7 cells, and COS-1 cells transfected with wild-type or mutant ER alpha, stimulated with EGF and E2, using Phospho-Estrogen Receptor alpha (Ser118) (16J4) Mouse mAb (upper) or control estrogen receptor alpha antibody #2512 (lower).
Estrogen Receptor α Activation Antibody Sampler Kit: Image 2
Immunohistochemical staining of phosphorylated estrogen receptor alpha in paraffin-embedded human breast carcinoma showing nuclear localization, using Phospho-Estrogen Receptor alpha (Ser118) (16J4) Mouse mAb.
Estrogen Receptor α Activation Antibody Sampler Kit: Image 3
Western blot analysis of extracts from MCF7 cells, either untreated (-), or treated (+) with combinations of the following treatments as indicated: Human Epidermal Growth Factor (hEGF) #8916 (100ng/ml, 15 min), or hEGF-treated and subsequently treated with calf intestinal phosphatase (CIP) and λ-phosphatase, using Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb (upper) or Estrogen Receptor α (D8H8) Rabbit mAb #8644 (lower).
Estrogen Receptor α Activation Antibody Sampler Kit: Image 4
Confocal immunofluorescent analysis of serum-starved MCF7 cells (ERα-positive), either untreated (upper left), stimulated with Human Epidermal Growth Factor (hEGF) #8916 (10 ng/mL, 30 min; upper right), or stimulated with hEGF and post-processed with λ-phosphatase (lower left), using Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb (green). hEGF-treated SK-BR-3 cells (ERα-negative; lower right) are included as an additional negative control. Actin filaments were labeled with DyLight™ 554 Phalloidin #13054 (red).
Estrogen Receptor α Activation Antibody Sampler Kit: Image 5
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Estrogen Receptor α Activation Antibody Sampler Kit: Image 6
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO* is added and emits light during enzyme catalyzed decomposition.
Estrogen Receptor α Activation Antibody Sampler Kit: Image 7
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and Estrogen Receptor α (D8H8) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across TFF1/pS2, a known target gene of Estrogen Receptor α (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product data sheet.
Estrogen Receptor α Activation Antibody Sampler Kit: Image 8
Western blot analysis of extracts from ER-positive cell lines (MCF7, T-47D, ZR-75-1) and ER-negative cell lines (SK-BR-3 and MCF 10A) using Estrogen Receptor α (D8H8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Estrogen Receptor α Activation Antibody Sampler Kit: Image 9
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and 5 μl of Estrogen Receptor α (D8H8) Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using SimpleChIP® ChIP-seq DNA Library Prep Kit for Illumina® #56795. The figure shows binding across chromosome 21 (upper), including TFF1/pS2 (lower), a known target gene of Estrogen Receptor α (see additional figure containing ChIP-qPCR data).
Estrogen Receptor α Activation Antibody Sampler Kit: Image 10
Chromatin immunoprecipitations were performed with cross-linked chromatin from MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 45 minutes and either Estrogen Receptor α (D8H8) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
To Purchase # 49757T
Product # Size Price
49757T
1 Kit  (3 x 20 microliters) $ 268

Product Description

The Estrogen Receptor α Activation Antibody Sampler Kit provides an economical means of detecting the activation of estrogen receptor α using phospho-specific and control antibodies. This kit includes enough antibodies to perform two western blot experiments with each primary antibody.

Specificity / Sensitivity

Estrogen Receptor α (D8H8) Rabbit mAb recognizes endogenous levels of total ERα protein. Phospho-Estrogen Receptor α (Ser118) (16J4) Mouse mAb detects endogenous levels of ERα protein only when phosphorylated at Ser118. It does not cross-react with phosphorylated estrogen receptor β. Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb recognizes endogenous levels of ERα protein only when phosphorylated at Ser167.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy terminus of human ERα protein. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser118 of human ERα protein. Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser167 of human ERα protein.

Background

Estrogen receptor α (ERα), a member of the steroid receptor superfamily, contains highly conserved DNA binding and ligand binding domains (1). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ERα regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (2). Phosphorylation at multiple sites provides an important mechanism to regulate ERα activity (3-5). Ser104, 106, 118, and 167 are located in the amino-terminal transcription activation function domain AF-1, and phosphorylation of these serine residues plays an important role in regulating ERα activity. Ser118 may be the substrate of the transcription regulatory kinase CDK7 (5). Ser167 may be phosphorylated by p90RSK and Akt (4,6). According to the research literature, phosphorylation at Ser167 may confer tamoxifen resistance in breast cancer patients (4).
  1. Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
  2. Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
  3. Chen, D. et al. (1999) Mol Cell Biol 19, 1002-15.
  4. Campbell, R.A. et al. (2001) J Biol Chem 276, 9817-24.
  5. Chen, D. et al. (2000) Mol Cell 6, 127-37.
  6. Joel, P.B. et al. (1998) Mol Cell Biol 18, 1978-84.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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For Research Use Only. Not For Use In Diagnostic Procedures.
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