Product: Estrogen Receptor-alpha Antibody
Catalog: AF6058
Description: Rabbit polyclonal antibody to Estrogen Receptor-alpha
Application: WB IHC IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Horse, Rabbit, Dog, Chicken, Xenopus
Mol.Wt.: 66kDa,55kDa; 66kD(Calculated).
Uniprot: P03372
RRID: AB_2834976

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(92%), Bovine(100%), Horse(100%), Rabbit(100%), Dog(100%), Chicken(100%), Xenopus(100%)
Clonality:
Polyclonal
Specificity:
Estrogen Receptor-alpha Antibody detects endogenous levels of total Estrogen Receptor-alpha.
RRID:
AB_2834976
Cite Format: Affinity Biosciences Cat# AF6058, RRID:AB_2834976.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

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;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P03372 ESR1_HUMAN:

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

Description:
ER-alpha is a nuclear hormone receptor and transcription factor. Regulates gene expression and affects cellular proliferation and differentiation in target tissues. Two splice-variant isoforms have been described.
Sequence:
MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Horse
100
Bovine
100
Dog
100
Xenopus
100
Chicken
100
Rabbit
100
Zebrafish
92
Sheep
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P03372 As Substrate

Site PTM Type Enzyme
T4 Phosphorylation
T7 Phosphorylation
S10 Phosphorylation
K32 Acetylation
S46 Phosphorylation
S47 Phosphorylation
Y52 Phosphorylation P00519 (ABL1)
S102 Phosphorylation P49841 (GSK3B)
S104 Phosphorylation P49841 (GSK3B) , P27361 (MAPK3) , P28482 (MAPK1) , P31749 (AKT1) , P24941 (CDK2) , Q00526 (CDK3) , P42345 (MTOR)
S106 Phosphorylation P24941 (CDK2) , Q00526 (CDK3) , P49841 (GSK3B) , P28482 (MAPK1) , P27361 (MAPK3) , P42345 (MTOR) , P31749 (AKT1)
S118 Phosphorylation P24941 (CDK2) , P27361 (MAPK3) , O15111 (CHUK) , P49841 (GSK3B) , P31749 (AKT1) , Q16539 (MAPK14) , P50613 (CDK7) , Q00526 (CDK3) , P28482 (MAPK1)
S154 Phosphorylation
S167 Phosphorylation O14965 (AURKA) , P68400 (CSNK2A1) , P51812 (RPS6KA3) , P31751 (AKT2) , P23443 (RPS6KB1) , Q14164 (IKBKE) , P31749 (AKT1) , Q15418 (RPS6KA1)
K171 Acetylation
K171 Methylation
K171 Sumoylation
K180 Methylation
K180 Sumoylation
S212 Phosphorylation
Y219 Phosphorylation P00519 (ABL1)
S236 Phosphorylation P17612 (PRKACA)
K244 Acetylation
K252 Acetylation
R260 Methylation
K266 Acetylation
K266 Methylation
K266 Sumoylation
K268 Acetylation
K268 Sumoylation
R277 Methylation
S282 Phosphorylation P68400 (CSNK2A1)
S294 Phosphorylation P24941 (CDK2) , Q16539 (MAPK14)
K299 Acetylation
K299 Sumoylation
K302 Acetylation
K302 Methylation
K302 Ubiquitination
K303 Acetylation
S305 Phosphorylation Q13153 (PAK1) , P31749 (AKT1) , P17612 (PRKACA) , O14965 (AURKA) , Q9UHD2 (TBK1) , O96013 (PAK4)
T311 Phosphorylation Q16539 (MAPK14)
S341 Phosphorylation
K401 Methylation
K472 Acetylation
K472 Sumoylation
T485 Phosphorylation
Y537 Phosphorylation P06239 (LCK) , P12931 (SRC)
S554 Phosphorylation
S559 Phosphorylation P68400 (CSNK2A1)
S576 Phosphorylation
S578 Phosphorylation
T594 Phosphorylation

Research Backgrounds

Function:

Nuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Ligand-dependent nuclear transactivation involves either direct homodimer binding to a palindromic estrogen response element (ERE) sequence or association with other DNA-binding transcription factors, such as AP-1/c-Jun, c-Fos, ATF-2, Sp1 and Sp3, to mediate ERE-independent signaling. Ligand binding induces a conformational change allowing subsequent or combinatorial association with multiprotein coactivator complexes through LXXLL motifs of their respective components. Mutual transrepression occurs between the estrogen receptor (ER) and NF-kappa-B in a cell-type specific manner. Decreases NF-kappa-B DNA-binding activity and inhibits NF-kappa-B-mediated transcription from the IL6 promoter and displace RELA/p65 and associated coregulators from the promoter. Recruited to the NF-kappa-B response element of the CCL2 and IL8 promoters and can displace CREBBP. Present with NF-kappa-B components RELA/p65 and NFKB1/p50 on ERE sequences. Can also act synergistically with NF-kappa-B to activate transcription involving respective recruitment adjacent response elements; the function involves CREBBP. Can activate the transcriptional activity of TFF1. Also mediates membrane-initiated estrogen signaling involving various kinase cascades. Isoform 3 is involved in activation of NOS3 and endothelial nitric oxide production. Isoforms lacking one or several functional domains are thought to modulate transcriptional activity by competitive ligand or DNA binding and/or heterodimerization with the full-length receptor. Essential for MTA1-mediated transcriptional regulation of BRCA1 and BCAS3. Isoform 3 can bind to ERE and inhibit isoform 1.

PTMs:

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>Peripheral membrane protein>Cytoplasmic side.
Note: A minor fraction is associated with the inner membrane.

Nucleus. Cytoplasm. Cell membrane>Peripheral membrane protein>Cytoplasmic side. Cell membrane>Single-pass type I membrane protein.
Note: Associated with the inner membrane via palmitoylation (Probable). At least a subset exists as a transmembrane protein with a N-terminal extracellular domain.

Nucleus. Golgi apparatus. Cell membrane.
Note: Colocalizes with ZDHHC7 and ZDHHC21 in the Golgi apparatus where most probably palmitoylation occurs. Associated with the plasma membrane when palmitoylated.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

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

Subunit Structure:

Binds DNA as a homodimer. Can form a heterodimer with ESR2. Isoform 3 can probably homodimerize or heterodimerize with isoform 1 and ESR2. Interacts with FOXC2, MAP1S, SLC30A9, UBE1C and NCOA3 coactivator (By similarity). Interacts with EP300; the interaction is estrogen-dependent and enhanced by CITED1. Interacts with CITED1; the interaction is estrogen-dependent. Interacts with NCOA5 and NCOA6 coactivators. Interacts with NCOA7; the interaction is a ligand-inducible. Interacts with PHB2, PELP1 and UBE1C. Interacts with AKAP13. Interacts with CUEDC2. Interacts with KDM5A. Interacts with SMARD1. Interacts with HEXIM1. Interacts with PBXIP1. Interaction with MUC1 is stimulated by 7 beta-estradiol (E2) and enhances ERS1-mediated transcription. Interacts with DNTTIP2, FAM120B and UIMC1. Interacts with isoform 4 of TXNRD1. Interacts with KMT2D/MLL2. Interacts with ATAD2 and this interaction is enhanced by estradiol. Interacts with KIF18A and LDB1. Interacts with RLIM (via C-terminus). Interacts with MACROD1. Interacts with SH2D4A and PLCG. Interaction with SH2D4A blocks binding to PLCG and inhibits estrogen-induced cell proliferation. Interacts with DYNLL1. Interacts with CCDC62 in the presence of estradiol/E2; this interaction seems to enhance the transcription of target genes. Interacts with NR2C1; the interaction prevents homodimerization of ESR1 and suppresses its transcriptional activity and cell growth. Interacts with DNAAF4. Interacts with PRMT2. Interacts with PI3KR1 or PI3KR2, SRC and PTK2/FAK1. Interacts with RBFOX2. Interacts with STK3/MST2 only in the presence of SAV1 and vice-versa. Binds to CSNK1D. Interacts with NCOA2; NCOA2 can interact with ESR1 AF-1 and AF-2 domains simultaneously and mediate their transcriptional synergy. Interacts with DDX5. Interacts with NCOA1; the interaction seems to require a self-association of N-terminal and C-terminal regions. Interacts with ZNF366, DDX17, NFKB1, RELA, SP1 and SP3. Interacts with NRIP1 (By similarity). Interacts with GPER1; the interaction occurs in an estrogen-dependent manner. Interacts with CLOCK and the interaction is stimulated by estrogen. Interacts with BCAS3. Interacts with TRIP4 (ufmylated); estrogen dependent. Interacts with LMTK3; the interaction phosphorylates ESR1 (in vitro) and protects it against proteasomal degradation. Interacts with CCAR2 (via N-terminus) in a ligand-independent manner. Interacts with ZFHX3. Interacts with SFR1 in a ligand-dependent and -independent manner. Interacts with DCAF13, LATS1 and DCAF1; regulates ESR1 ubiquitination and ubiquitin-mediated proteasomal degradation. Interacts (via DNA-binding domain) with POU4F2 (C-terminus); this interaction increases the estrogen receptor ESR1 transcriptional activity in a DNA- and ligand 17-beta-estradiol-independent manner (By similarity). Interacts with ESRRB isoform 1. Interacts with UBE3A and WBP2. Interacts with GTF2B. Interacts with RBM39 (By similarity). In the absence of hormonal ligand, interacts with TACC1.

Family&Domains:

Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain. The modulating domain, also known as A/B or AF-1 domain has a ligand-independent transactivation function. The C-terminus contains a ligand-dependent transactivation domain, also known as E/F or AF-2 domain which overlaps with the ligand binding domain. AF-1 and AF-2 activate transcription independently and synergistically and act in a promoter- and cell-specific manner. AF-1 seems to provide the major transactivation function in differentiated cells.

Belongs to the nuclear hormone receptor family. NR3 subfamily.

Research Fields

· Human Diseases > Drug resistance: Antineoplastic > Endocrine resistance.

· Human Diseases > Cancers: Overview > Pathways in cancer.   (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Breast cancer.   (View pathway)

· Organismal Systems > Endocrine system > Estrogen signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Prolactin signaling pathway.   (View pathway)

· Organismal Systems > Endocrine system > Thyroid hormone signaling pathway.   (View pathway)

· Organismal Systems > Excretory system > Endocrine and other factor-regulated calcium reabsorption.

References

1). Combined effects of di (2-ethylhexyl) phthalate and bisphenol A on thyroid hormone homeostasis in adolescent female rats. Environmental science and pollution research international, 2020 (PubMed: 32681327) [IF=5.8]

Application: IHC    Species: rat    Sample: thyroid

Fig. 5| Effects of DEHP and BPA on relative gene expression in the thyroid (n = 7). a Immunohistochemistry for Esr1 in the thyroid(original magnification, × 20). Scale bar = 200 μm. b The average optical density of Esr1 in the thyroid. c The mRNA expression of Esr1.d Immunohistochemistry for phosphorylation of Creb in the thyroid(original magnification, × 20).

2). Alleviation of Fufang Fanshiliu decoction on type II diabetes mellitus by reducing insulin resistance: A comprehensive network prediction and experimental validation. Journal of Ethnopharmacology, 2022 (PubMed: 35568115) [IF=5.4]

Application: WB    Species: Human    Sample: HepG2 cells

Fig. 8. FFSLD alleviated IR in the HepG2-IR cells model by regulating the PI3K-AKT and ESR pathways. (A, C) Western Blot detection and (B, D) quantitative analysis for the expressions of p-AKT, ESR1 in HepG2-IR cells treated with different concentrations of FFSLD for 24 h. Data (n = 3) are expressed as mean ± SEM. *p < 0.05, comparing with control.

3). Mechanism of Astragalus membranaceus (Huangqi, HQ) for treatment of heart failure based on network pharmacology and molecular docking. Journal of cellular and molecular medicine, 2024 (PubMed: 38780500) [IF=5.3]

Application: WB    Species: Human    Sample:

FIGURE 7 Effects of five active compounds on ESR1 level in Ang II‐induced cardiomyocytes. Cardiomyocytes were treated with Ang II (1 μM) for 24 h in the presence or absence of Quercetin (20 μM), Isorhamnetin (50 μM), Calycosin (20 μM), Kaempferol (20 μM), or Formononetin (20 μM) and then examined for the ESR1 level in cardiomyocytes by ELISA assay (A); the protein levels of ESR1 and phosphorylated‐ESR1 in cardiomyocytes by western blot assay (B). N = 3 (biological replicates). ***p 

4). The Role of Estrogen Receptor α in Response to Longitudinal Bone Growth in ob/ob Mice. Frontiers in Endocrinology, 2021 (PubMed: 34925230) [IF=5.2]

Application: WB    Species: Mice    Sample: femoral and spine

Figure 5 The expression level of estrogen receptors (ERs) in femur and spine. (A) Western blotting results of expression level of ERα and ERβ in femoral and spine growth plate (GP), n = 3. (B–E) Semiquantitative analysis of ER expression levels. (F) Quantitative real-time PCR results of ER genes in femoral and vertebral GP, n = 4. (G–J) Immunohistochemistry (IHC) results of expression level of ERα and ERβ in femoral and vertebral GP, n = 6. Scale bar: 20 μm. (K) Comparison of ERα HSCORE between femoral and spine GP (FGP, femoral growth plate; SGP, spine growth plate). (L, M) Results of Spearman’s analysis between ERα HSCORE and femur/spine length. *p < 0.05, ***p < 0.001 and ****p < 0.0001; ns denotes not significant.

Application: IHC    Species: Mice    Sample: femoral and vertebral

Figure 5 The expression level of estrogen receptors (ERs) in femur and spine. (A) Western blotting results of expression level of ERα and ERβ in femoral and spine growth plate (GP), n = 3. (B–E) Semiquantitative analysis of ER expression levels. (F) Quantitative real-time PCR results of ER genes in femoral and vertebral GP, n = 4. (G–J) Immunohistochemistry (IHC) results of expression level of ERα and ERβ in femoral and vertebral GP, n = 6. Scale bar: 20 μm. (K) Comparison of ERα HSCORE between femoral and spine GP (FGP, femoral growth plate; SGP, spine growth plate). (L, M) Results of Spearman’s analysis between ERα HSCORE and femur/spine length. *p < 0.05, ***p < 0.001 and ****p < 0.0001; ns denotes not significant.

5). Ormeloxifene, a selective estrogen receptor modulator, protects against pulmonary hypertension. European Journal of Pharmacology, 2023 (PubMed: 36731722) [IF=5.0]

6). Estrogen receptors and hypoxia inducible factor 1 alpha expression in abdominal wall endometriosis. REPRODUCTIVE BIOMEDICINE ONLINE, 2020 (PubMed: 32444257) [IF=4.0]

Application: IHC    Species: human    Sample: endometrium

FIGURE 1| Expression and localization of oestrogen receptors and hypoxia-inducible factor-1alpha (HIF-1a) proteins in proliferative phase and secretary phase of normal control endometrium analysed by immunohistochemistry. (A–D) ERa; (E–H) ERb; (I–L) G protein-coupled oestrogen receptor (GPER); (M–P) HIF-1a. Micrographs were taken at magnifications of × 200 (left panels) and marked areas in left-hand panels shown at × 400 (right panels) respectively. ER, oestrogen receptor.

7). Anti‑chondrocyte apoptosis effect of genistein in treating inflammation‑induced osteoarthritis. Molecular Medicine Reports, 2020 (PubMed: 32582961) [IF=3.4]

Application: WB    Species: human    Sample: chondrocytes

Figure 2. |Protein expression levels of collagen II and aggrecan increase with the added concentration of genistein, while caspase 3 protein levels gradually decrease in each group. (A) Western blot analysis of samples from the experimental groups were tested with the indicated antibodies.

8). Effects of l-arginine on endometrial estrogen receptor α/β and progesterone receptor expression in nutrient-restricted sheep. THERIOGENOLOGY, 2019 (PubMed: 31352175) [IF=2.8]

Application: WB    Species: sheep    Sample: endometrium

Fig. 2. | Location and expression of ERa in the ovine endometrium. A) Immunohistochemical localization of ERa protein in the ovine uterus; legend: LE luminal epithelium, ES endometrial stroma, GE glandular epithelium, Myo myometrium, Bar ¼ 100 mm. A1) Negative control (no primary antibody). B) relative expression of ESR1 mRNA. C) Location of ERa and b-actin protein following Western blot.

Application: IHC    Species: sheep    Sample: uterus

Fig. 2. | Location and expression of ERa in the ovine endometrium. A) Immunohistochemical localization of ERa protein in the ovine uterus; legend: LE luminal epithelium, ES endometrial stroma, GE glandular epithelium, Myo myometrium, Bar ¼ 100 mm. A1) Negative control (no primary antibody). B) relative expression of ESR1 mRNA. C) Location of ERa and b-actin protein following Western blot.

9). Establishment of a New Cell Line of Canine Inflammatory Mammary Cancer: IMC‐118. Veterinary and Comparative Oncology, 2022 (PubMed: 35429113) [IF=2.1]

10). Difference in Expressional Profile of Compact and Expanded-Type Equine Cumulus-Oocyte-Complexes. , 2022

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