Phospho-Estrogen Receptor alpha (S118) Recombinant Rabbit Monoclonal Antibody [SC05-87]
cat.: ET1610-32
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IF-Tissue, IHC-P
Clonality: Monoclonal
Clone number: SC05-87
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 66 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser118 of Human ESR1 aa 110-154 / 595.
Positive control: MCF7 whole cell lysate, MCF7 treated with 100nM Estradiol for 30 minutes add 100ng/ml EGF for 5 minutes whole cell lysate, MCF7 treated with 100nM Estradiol for 30 minutes then treated with 100ng/ml EGF for 5 minutes whole cell lysate, Hela, MCF-7, HUVEC, human breast carcinoma tissue, human breast tissue, mouse placenta tissue, human placenta tissue, human fallopian tube tissue, human uterus tissue.
Subcellular location: Nucleus, Cytoplasm, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
1:1,000
1:50-1:200
1:50-1:200
1:50-1:500
Uniprot #: SwissProt: P03372 Human | P19785 Mouse
Alternative names: DKFZp686N23123 ER alpha ER ER-alpha Era ESR ESR1 ESR1_HUMAN ESRA Estradiol receptor Estrogen nuclear receptor alpha Estrogen receptor 1 Estrogen receptor alpha 3*,4,5,6,7*/822 isoform Estrogen receptor alpha Estrogen receptor alpha delta 3*,4,5,6,7*,8*/941 isoform Estrogen receptor alpha delta 3*,4,5,6,7*/819 2 isoform Estrogen receptor alpha delta 4 +49 isoform Estrogen receptor alpha delta 4*,5,6,7*/654 isoform Estrogen receptor alpha delta 4*,5,6,7,8*/901 isoform Estrogen receptor alpha E1 E2 1 2 Estrogen receptor alpha E1 N2 E2 1 2 Estrogen receptor ESTRR NR3A1 Nuclear receptor subfamily 3 group A member 1
Images
ET1610-32_1.jpg Fig1: Western blot analysis of Phospho-Estrogen Receptor alpha (S118) on different lysates with Rabbit anti-Phospho-Estrogen Receptor alpha (S118) antibody (ET1610-32) at 1/1,000 dilution.

Lane 1: MCF7 whole cell lysate
Lane 2: MCF7 treated with 100nM Estradiol for 30 minutes add 100ng/ml EGF for 5 minutes whole cell lysate
Lane 3: MCF7 treated with 100nM Estradiol for 30 minutes then treated with 100ng/ml EGF for 5 minutes whole cell lysate
Lane 4: MCF7 whole cell lysate treated with λpp for 1 hour
Lane 5: MCF7 treated with 100nM Estradiol for 30 minutes add 100ng/ml EGF for 5 minutes whole cell lysate treated with λpp for 1 hour
Lane 6: MCF7 treated with 100nM Estradiol for 30 minutes then treated with 100ng/ml EGF for 5 minutes whole cell lysate treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 66 kDa
Observed band size: 66 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-32) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
ET1610-32_2.jpg Fig2: ICC staining of Phospho-Estrogen Receptor alpha (S118) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-32_3.jpg Fig3: ICC staining of Phospho-Estrogen Receptor alpha (S118) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-32_4.jpg Fig4: ICC staining of Phospho-Estrogen Receptor alpha (S118) in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-32, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1610-32_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-32_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-32_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse placenta tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-32_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-32_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human fallopian tube tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1610-32_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Phospho-Estrogen Receptor alpha (S118) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-32, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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