重组Anti-STAT3 (phospho S727)抗体[E121-31] (ab32143)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E121-31] to STAT3 (phospho S727)
- Suitable for: ChIC/CUT&RUN-seq, ICC/IF, WB, Dot blot, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
-
产品名称
Anti-STAT3 (phospho S727)抗体[E121-31]
参阅全部 STAT3 一抗 -
描述
兔单克隆抗体[E121-31] to STAT3 (phospho S727) -
宿主
Rabbit -
经测试应用
适用于: ChIC/CUT&RUN-seq, ICC/IF, WB, Dot blot, IP, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Horse, Cow, Macaque monkey -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: A431 cell lysate, C6 treated with epidermal growth factor. IP: HeLa cells ICC/IF: A431 cells IHC-P: human astrocytoma, rat cerebral cortex, mouse liver, and brain astrocytoma tissues ChIC/CUT&RUN seq: HepG2 cell
-
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: We have preliminary internal testing data to indicate this antibody may not react with this 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. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E121-31 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Assay kits
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32143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.
|
|
ICC/IF |
1/500.
For unpurified, use 1/100. |
|
WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 88 kDa).
Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls. |
Dot blot |
1/1000.
|
|
IP |
1/60.
|
|
IHC-P | (5) |
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified, use 1/50. |
说明 |
---|
ChIC/CUT&RUN-seq
Use at an assay dependent concentration. |
ICC/IF
1/500. For unpurified, use 1/100. |
WB
1/1000 - 1/10000. Detects a band of approximately 98 kDa (predicted molecular weight: 88 kDa). Stimulation may be required to allow detection of the phosphorylated protein. Please see images below for recommended treatment conditions and positive controls. |
Dot blot
1/1000. |
IP
1/60. |
IHC-P
1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified, use 1/50. |
靶标
-
功能
Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. -
组织特异性
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. -
疾病相关
Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
Autoimmune disease, multisystem, infantile-onset -
序列相似性
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
翻译后修饰
Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling. -
细胞定位
Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1. - Information by UniProt
-
数据库链接
- Entrez Gene: 508541 Cow
- Entrez Gene: 6774 Human
- Entrez Gene: 20848 Mouse
- Entrez Gene: 25125 Rat
- Omim: 102582 Human
- SwissProt: P61635 Cow
- SwissProt: P40763 Human
- SwissProt: P42227 Mouse
see all -
别名
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
see all
图片
-
ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 cells (starved overnight and treated with 100ng/ml IL-6 for 30min) and 5 µg of ab32143 [E121-31]. 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.
Additional screenshots of mapped reads can be downloaded here. The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods. -
Purified ab32143 staining STAT3 (phospho S727) in A431 cells by Immunocytochemistry/ Immunofluorescence. 4% PFA-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma) cells labelled with ab32143 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing nuclear staining on A431 cell line. The red staining is ab7291 anti-Tubulin (mouse mAb), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)Immunohistochemical staining of paraffin embedded human astrocytoma with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
-
All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/5000 dilution (purified)
Lane 1 : C6 treated with epidermal growth factor
Lane 2 : untreated C6 whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
All lanes : purified
Lane 1 : A431 treated with epidermal growth factor
Lane 2 : untreated A431 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)Immunohistochemical staining of paraffin embedded rat cerebral cortex with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)Immunohistochemical staining of paraffin embedded mouse liver with purified ab32143 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
-
Immunocytochemical/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling STAT3 (phospho S727) with ab32143 (left) and STAT3 with ab32500 (right) both at a dilution of 1/500.
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) (1/1000) was used as the secondary antibody (green). DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (1/200) was used as a counterstain (red).
The green staining was increased and translocated from the cytoplasm into the nucleus in the EGF (ab9697 100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signal was decreased. For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment.
-
Dot Blot analysis of Lane 1: STAT3 (pS727) phospho peptide and Lane 2: STAT3 non-phospho peptide labeling STAT3 (phospho S727) with ab32143 at 1/1000 dilution (0.009 μg/ml). 5% NFDM /TBST was used as the diluting and blocking buffer and concentration. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100,000 dilution. Exposure time: 10 seconds.
-
All lanes : Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143) at 1/1000 dilution (unpurified)
Lane 1 : A431 cell lysate
Lane 2 : A431 + EGF cell lysate
Predicted band size: 88 kDa
Observed band size: 98 kDa why is the actual band size different from the predicted? -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT3 (phospho S727) antibody [E121-31] (ab32143)
IHC-P analysis of brain astrocytoma using unpurified ab32143 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
-
ab32143 (purified) at 1/60 dilution (2.594 µg/ml) immunoprecipitating STAT3 in HeLa whole cell lysate.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab32143 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32143 in HeLa whole cell lysateFor western blotting, ab32143 at 1/500 and VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST.
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 (89)
ab32143 被引用在 89 文献中.
- Cai M et al. Fibroblast Growth Factor 21 Relieves Lipopolysaccharide-Induced Acute Lung Injury by Suppressing JAK2/STAT3 Signaling Pathway. Inflammation 47:209-226 (2024). PubMed: 37864659
- Luan Y et al. Artesunate regulates the proliferation and differentiation of neural stem cells by activating the JAK‑2/STAT‑3 signaling pathway in ischemic stroke. Exp Ther Med 25:2 (2023). PubMed: 36561626
- Chen T et al. Splicing factor SF3B3, a NS5-binding protein, restricts ZIKV infection by targeting GCH1. Virol Sin 38:222-232 (2023). PubMed: 36572150
- Reicherz A et al. Indwelling stents cause severe inflammation and fibrosis of the ureter via urothelial-mesenchymal transition. Sci Rep 13:5492 (2023). PubMed: 37015949
- Xu L et al. 4-Octyl itaconate attenuates LPS-induced acute kidney injury by activating Nrf2 and inhibiting STAT3 signaling. Mol Med 29:58 (2023). PubMed: 37095432