重组Anti-Estrogen Receptor alpha (phospho S118)抗体[E91] (ab32396)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E91] to Estrogen Receptor alpha (phospho S118)
- Suitable for: ChIC/CUT&RUN-seq, Dot blot, WB, ICC/IF
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-Estrogen Receptor alpha (phospho S118)抗体[E91]
参阅全部 Estrogen Receptor alpha 一抗 -
描述
兔单克隆抗体[E91] to Estrogen Receptor alpha (phospho S118) -
宿主
Rabbit -
特异性
This antibody only detects ER alpha phosphorylated on Serine 118.
-
经测试应用
适用于: ChIC/CUT&RUN-seq, Dot blot, WB, ICC/IFmore details
不适用于: Flow Cyt or IHC-P -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Estrogen Receptor alpha. The exact sequence is proprietary.
-
阳性对照
- MCF7 cell lysate human breast adenocarcinoma
-
常规说明
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E91 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Assay kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32396于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
---|---|---|
ChIC/CUT&RUN-seq |
Use at an assay dependent concentration.
|
|
Dot blot |
1/1000.
|
|
WB | (2) |
1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
|
ICC/IF |
1/200.
For unpurified use at 1/50 - 1/100 dilution. |
说明 |
---|
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
Dot blot
1/1000. |
WB
1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa). |
ICC/IF
1/200. For unpurified use at 1/50 - 1/100 dilution. |
靶标
-
功能
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. Can activate the transcriptional activity of TFF1. -
序列相似性
Belongs to the nuclear hormone receptor family. NR3 subfamily.
Contains 1 nuclear receptor DNA-binding domain. -
结构域
Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain. -
翻译后修饰
Phosphorylated by cyclin A/CDK2. Phosphorylation probably enhances transcriptional activity.
Glycosylated; contains N-acetylglucosamine, probably O-linked.
Ubiquitinated. Deubiquitinated by OTUB1.
Dimethylated by PRMT1 at Arg-260. The methylation may favor cytoplasmic localization.
Palmitoylated (isoform 3). Not biotinylated (isoform 3). -
细胞定位
Nucleus. Cytoplasm. Cell membrane. A minor fraction is associated with the inner membrane and Nucleus. Cytoplasm. Cell membrane. Associated with the inner membrane via palmitoylation. - Information by UniProt
-
数据库链接
- Entrez Gene: 2099 Human
- Omim: 133430 Human
- SwissProt: P03372 Human
- Unigene: 208124 Human
-
别名
- 7*/654 isoform antibody
- 7*/819 2 isoform antibody
- 7*/822 isoform antibody
see all
图片
-
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/µL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
-
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/µL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
-
ChIC/CUT&RUN was performed using a pAG-MNase at a final concentration of 700 ng/µL, 2.5 x 10^5 MCF7 (Human breast adenocarcinoma epithelial cell) cells treated with phenol red free medium and 5% charcoal stripped FBS for 3 days than treated with β-estradiol (10 nM 45 min) and 5 µg of ab32396 [E91]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.
-
Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) treated with EGF (100ng/ml, 5min) and treated with Lambda Protein Phosphatase 31℃ for 2h cells labeling Estrogen receptor alpha (phospho S118) with purified ab32396 at 1:200 dilution (8.9μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
-
All lanes : Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) at 1/1000 dilution (purified)
Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates
Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 66 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
Exposure time: 5 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
-
Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 5 μg/ml (1/200 dilution). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, an AlexaFluor®488 Goat anti-Rabbit was used as the secondary antibody at 2 μg/ml (1/1000 dilution). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 2.5 μg/ml (1/200 dilution). DAPI nuclear counterstain.
Confocal image showing the signal increased after EGF (100ng/ml, 5 min) treatment and decreased after Lambda Protein Phosphatase treatment (31°C for 2 hours). -
Dot blot analysis of Lane 1: Estrogen Receptor alpha (pS118) phospho peptide and Lane 2: Estrogen Receptor alpha non-phospho peptide labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.
-
Immunofluorescent staining of (A) untreated and (B) Phosphatase-treated MCF-7 cells using unpurified ab32396.
-
All lanes : Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) at 1/1000 dilution (unpurified)
Lane 1 : MCF7 cell lysate (untreated).
Lane 2 : MCF7 cell lysate (treated with b-Estradiol and EGF).
Predicted band size: 66 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
-
SDS download
-
Datasheet download
文献 (18)
ab32396 被引用在 18 文献中.
- Baumgartner NE et al. History of Previous Midlife Estradiol Treatment Permanently Alters Interactions of Brain Insulin-like Growth Factor-1 Signaling and Hippocampal Estrogen Synthesis to Enhance Cognitive Aging in a Rat Model of Menopause. J Neurosci 42:7969-7983 (2022). PubMed: 36261268
- Zhao Z et al. Crosstalk between the muscular estrogen receptor a and BDNF/TrkB signaling alleviates metabolic syndrome via 7,8-dihydroxyflavone in female mice. Mol Metab 45:101149 (2021). PubMed: 33352311
- Ohlsson C et al. Phosphorylation site S122 in estrogen receptor a has a tissue-dependent role in female mice. FASEB J 34:15991-16002 (2020). PubMed: 33067917
- Ma Y et al. Loss of Estrogen Efficacy Against Hippocampus Damage in Long-Term OVX Mice Is Related to the Reduction of Hippocampus Local Estrogen Production and Estrogen Receptor Degradation. Mol Neurobiol 57:3540-3551 (2020). PubMed: 32542593
- Castellaro AM et al. Tumor-Associated Macrophages Induce Endocrine Therapy Resistance in ER+ Breast Cancer Cells. Cancers (Basel) 11:N/A (2019). PubMed: 30736340