重组Anti-EGFR (phospho Y845)抗体[EPR2149Y] (ab97613)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2149Y] to EGFR (phospho Y845)
- Suitable for: Flow Cyt (Intra), Dot blot, WB, ICC/IF
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-EGFR (phospho Y845)抗体[EPR2149Y]
参阅全部 EGFR 一抗 -
描述
兔单克隆抗体[EPR2149Y] to EGFR (phospho Y845) -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), Dot blot, WB, ICC/IFmore details
不适用于: IHC-P -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human EGFR (phospho S845). The exact sequence is proprietary.
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阳性对照
- A431 cells and cell lysates.
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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
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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, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2149Y -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab97613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 |
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Flow Cyt (Intra) |
1/400.
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Dot blot |
Use at an assay dependent concentration.
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WB |
1/500. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa).
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ICC/IF |
1/100 - 1/250.
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说明 |
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Flow Cyt (Intra)
1/400. |
Dot blot
Use at an assay dependent concentration. |
WB
1/500. Detects a band of approximately 175 kDa (predicted molecular weight: 134 kDa). |
ICC/IF
1/100 - 1/250. |
靶标
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功能
Receptor tyrosine kinase binding ligands of the EGF family and activating several signaling cascades to convert extracellular cues into appropriate cellular responses. Known ligands include EGF, TGFA/TGF-alpha, amphiregulin, epigen/EPGN, BTC/betacellulin, epiregulin/EREG and HBEGF/heparin-binding EGF. Ligand binding triggers receptor homo- and/or heterodimerization and autophosphorylation on key cytoplasmic residues. The phosphorylated receptor recruits adapter proteins like GRB2 which in turn activates complex downstream signaling cascades. Activates at least 4 major downstream signaling cascades including the RAS-RAF-MEK-ERK, PI3 kinase-AKT, PLCgamma-PKC and STATs modules. May also activate the NF-kappa-B signaling cascade. Also directly phosphorylates other proteins like RGS16, activating its GTPase activity and probably coupling the EGF receptor signaling to the G protein-coupled receptor signaling. Also phosphorylates MUC1 and increases its interaction with SRC and CTNNB1/beta-catenin.
Isoform 2 may act as an antagonist of EGF action. -
组织特异性
Ubiquitously expressed. Isoform 2 is also expressed in ovarian cancers. -
疾病相关
Lung cancer
Inflammatory skin and bowel disease, neonatal, 2 -
序列相似性
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
翻译后修饰
Phosphorylation at Ser-695 is partial and occurs only if Thr-693 is phosphorylated. Phosphorylation at Thr-678 and Thr-693 by PRKD1 inhibits EGF-induced MAPK8/JNK1 activation. Dephosphorylation by PTPRJ prevents endocytosis and stabilizes the receptor at the plasma membrane. Autophosphorylation at Tyr-1197 is stimulated by methylation at Arg-1199 and enhances interaction with PTPN6. Autophosphorylation at Tyr-1092 and/or Tyr-1110 recruits STAT3. Dephosphorylated by PTPN1 and PTPN2.
Monoubiquitinated and polyubiquitinated upon EGF stimulation; which does not affect tyrosine kinase activity or signaling capacity but may play a role in lysosomal targeting. Polyubiquitin linkage is mainly through 'Lys-63', but linkage through 'Lys-48', 'Lys-11' and 'Lys-29' also occurs. Deubiquitination by OTUD7B prevents degradation. Ubiquitinated by RNF115 and RNF126.
Methylated. Methylation at Arg-1199 by PRMT5 stimulates phosphorylation at Tyr-1197. -
细胞定位
Secreted and Cell membrane. Endoplasmic reticulum membrane. Golgi apparatus membrane. Nucleus membrane. Endosome. Endosome membrane. Nucleus. In response to EGF, translocated from the cell membrane to the nucleus via Golgi and ER. Endocytosed upon activation by ligand. Colocalized with GPER1 in the nucleus of estrogen agonist-induced cancer-associated fibroblasts (CAF). - Information by UniProt
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数据库链接
- Entrez Gene: 1956 Human
- Omim: 131550 Human
- SwissProt: P00533 Human
- Unigene: 488293 Human
- Unigene: 605083 Human
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别名
- Avian erythroblastic leukemia viral (v erb b) oncogene homolog antibody
- Cell growth inhibiting protein 40 antibody
- Cell proliferation inducing protein 61 antibody
see all
图片
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All lanes : Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613) at 1/1500 dilution (purified)
Lane 1 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma cell line) treated with 100 ng/ml EGF for 10 minutes whole cell lysates
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: 134 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 10 minutes cells labeling EGFR with purified ab97613 at 1/400 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated A431 cells (Green).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed,0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cell lines labeling EGFR with ab97613 (purified) at 1/50 dilution,followed by Goat anti-Rabbit IgG H & L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution
Cells were counterstained with ab195889 (Anti-alpha Tubulin antibody [DM1A] ) - Microtubule Marker (Alexa Fluor® 594)
The nuclear counterstain is DAPI (blue).
The green staining on the membrane was increased in the EGF (100ng/ml, 10min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.
For the pan antibody, there was no great difference after EGF (100ng/ml, 10min) or EGF (100ng/ml, 10min) + LP treatment. The data showed mostly membranous staining.
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Dot Blot analysis of Lane 1: EGFR (pY845) phospho peptide and Lane 2: EGFR non-phospho peptide labeling EGFR (phospho Y845) with ab97613 (unpurified) 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.
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All lanes : Anti-EGFR (phospho Y845) antibody [EPR2149Y] (ab97613) at 1/500 dilution (unpurified)
Lane 1 : A431 cell lysates, untreated
Lane 2 : A431 cell lysates, treated with EGF
Lysates/proteins at 10 µg per lane.
Predicted band size: 134 kDa
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (4)
ab97613 被引用在 4 文献中.
- Komuro A et al. Identification of a novel fusion gene HMGA2-EGFR in glioblastoma. Int J Cancer 142:1627-1639 (2018). PubMed: 29193056
- Wang J et al. BIN1 reverses PD-L1-mediated immune escape by inactivating the c-MYC and EGFR/MAPK signaling pathways in non-small cell lung cancer. Oncogene 36:6235-6243 (2017). PubMed: 28714960
- Saghizadeh M et al. Adenovirus-driven overexpression of proteinases in organ-cultured normal human corneas leads to diabetic-like changes. Brain Res Bull 81:262-72 (2010). PubMed: 19828126
- Saghizadeh M et al. Normalization of wound healing and diabetic markers in organ cultured human diabetic corneas by adenoviral delivery of c-Met gene. Invest Ophthalmol Vis Sci 51:1970-80 (2010). PubMed: 19933191