PRODUCT CODE: ET1610-32

Recombinant Phospho Estrogen Receptor alpha (S118) Monoclonal Antibody (ET1610-32)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

Western blot analysis of Phospho-Estrogen Receptor alpha(S118) on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • Western blot analysis of Phospho-Estrogen Receptor alpha(S118) on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
  • 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 1% Blocker BSA 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 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • 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 1% Blocker BSA 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 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • 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 1% Blocker BSA 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 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • 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 Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% 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.
  • 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 Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% 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.
  • 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 Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% 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.
Western blot analysis of Phospho-Estrogen Receptor alpha(S118) on MCF-7 cell lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1610-32, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Recombinant Phospho Estrogen Receptor alpha (S118) Monoclonal Antibody (ET1610-32)

Immunogen

Recombinant protein

Host

Rabbit

Modification

Phospho

Modification Site

S118

Positive Control

MCF-7 cell lysates, Hela, MCF-7, HUVEC, human breast carcinoma tissue, human breast tissue, mouse placenta tissue, human placenta tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SC05-87

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

66 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-Estrogen Receptor alpha(S118)

SYNONYMS

DKFZp686N23123 antibody; ER alpha antibody; ER antibody; ER-alpha antibody; Era antibody; ESR antibody; ESR1 antibody; ESR1_HUMAN antibody; ESRA antibody; Estradiol receptor antibody; Estrogen nuclear receptor alpha antibody; Estrogen receptor 1 antibody; Estrogen receptor alpha 3*,4,5,6,7*/822 isoform antibody; Estrogen receptor alpha antibody; Estrogen receptor alpha delta 3*,4,5,6,7*,8*/941 isoform antibody; Estrogen receptor alpha delta 3*,4,5,6,7*/819 2 isoform antibody; Estrogen receptor alpha delta 4 +49 isoform antibody; Estrogen receptor alpha delta 4*,5,6,7*/654 isoform antibody; Estrogen receptor alpha delta 4*,5,6,7,8*/901 isoform antibody; Estrogen receptor alpha E1 E2 1 2 antibody; Estrogen receptor alpha E1 N2 E2 1 2 antibody; Estrogen receptor antibody; ESTRR antibody; NR3A1 antibody; Nuclear receptor subfamily 3 group A member 1 antibody

SEQUENCE SIMILARITIES

Belongs to the nuclear hormone receptor family. NR3 subfamily.

TISSUE SPECIFICITY

Widely expressed. Isoform 3 is not expressed in the pituitary gland.

POST-TRANSLATIONAL MODIFICATION

Phosphorylated by cyclin A/CDK2 and CK1. Phosphorylation probably enhances transcriptional activity. Self-association induces phosphorylation. Dephosphorylation at Ser-118 by PPP5C inhibits its transactivation activity. Phosphorylated by LMTK3 in vitro.; Glycosylated; contains N-acetylglucosamine, probably O-linked.; Ubiquitinated; regulated by LATS1 via DCAF1 it leads to ESR1 proteasomal degradation. Deubiquitinated by OTUB1.; Dimethylated by PRMT1 at Arg-260. The methylation may favor cytoplasmic localization. Demethylated by JMJD6 at Arg-260.; Palmitoylated (isoform 3). Not biotinylated (isoform 3).; Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation, but not for signaling mediated by the nuclear hormone receptor.

SUBCELLULAR LOCATION

Nucleus, Cytoplasm, Cell membrane.

FUNCTION

Estrogen receptor alpha (ERa, ER, ESR, ESRA, Era, NR3A1, estrogen receptor 1) is a ligand-activated transcription factor composed of several domains important for hormone binding, DNA binding and activation of transcription. Alternative splicing results in several ERa mRNA transcripts, which differ primarily in their 5' untranslated regions. ERa undergoes phosphorylation in response to estradiol binding. Human ERa is predominately phosphorylated on Ser 118 and to a lesser extent on Ser 104 and Ser 106. In response to activation of the mitogen-activated protein kinase pathway, phosphorylation occurs on Ser 118 and Ser 167. These serine residues are all located within the activation function 1 region of the N-terminal domain of ERa. In contrast, activation of protein kinase A increases the phosphorylation of Ser 236, which is located in the DNA-binding domain. Src kinase-dependent Tyr 537 phosphorylation may enhance estrogen binding to ERa. Mutation of Tyr 537 of the human ERa produces receptors having a range of constitutive activity.