重组Anti-EGFR (phospho Y1068)抗体[Y38] (ab32430)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y38] to EGFR (phospho Y1068)
- Suitable for: WB, ICC/IF, IP
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-EGFR (phospho Y1068)抗体[Y38]
参阅全部 EGFR 一抗 -
描述
兔单克隆抗体[Y38] to EGFR (phospho Y1068) -
宿主
Rabbit -
特异性
This antibody recognises only EGFR phosphorylated on Tyrosine 1068 of the mature human isoform 1 (corresponding to Y1092 from the precursor form P00533-1/p170).
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经测试应用
适用于: WB, ICC/IF, IPmore details
不适用于: Flow Cyt -
种属反应性
与反应: Human
预测可用于: Pig -
免疫原
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阳性对照
- WB: A431 cell lysate treated with EGF. ICC/IF: A431 cell lysate treated with EGF.
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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
Y38 -
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
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32430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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WB |
1/1000 - 1/10000. Predicted molecular weight: 135 kDa.
A blocking peptide is available for this antibody as ab205609. |
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ICC/IF |
1/100 - 1/250.
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IP |
1/30.
For unpurified format use at 1/150 dilution. |
说明 |
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WB
1/1000 - 1/10000. Predicted molecular weight: 135 kDa. A blocking peptide is available for this antibody as ab205609. |
ICC/IF
1/100 - 1/250. |
IP
1/30. For unpurified format use at 1/150 dilution. |
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
- Entrez Gene: 397070 Pig
- 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 Y1068) antibody [Y38] (ab32430) at 1/1000 dilution (Purified)
Lane 1 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates
Lane 2 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes whole cell lysates
Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) treated with 100 ng/ml EGF for 30 minutes. 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: 135 kDa
Observed band size: 175 kDa why is the actual band size different from the predicted? -
ab32430 (purified) at 1/30 dilution (2ug) immunoprecipitating EGFR in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug . A431 (Human epidermoid carcinoma epithelial cell) treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
Lane 2 (+): ab32430 & A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32430 in A431 treated with 200 ng/ml EGF for 15 minutes whole cell lysate 10ug
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
Immunocytochemistry/Immunofluorescence analysis of untreated, EGF treated and EGF + LP treated A431 cells labelling EGFR (phospho Y1068) with purified ab32430 (left) and EGFR with ab52894 (right) both at a dilution of 1/1000.
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
The image shows increased membrane staining after EGF (100ng/ml, 30min) treatment on A431 cells. The LP treatment decreased the increased membrane staining caused by EGF.
ab52894 was used as a Pan control for ab32430. The results showed membrane and cytoplasmic staining on untreated, EGF and EGF + LP treated A431 cells.
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
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SDS download
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Datasheet download
文献 (37)
ab32430 被引用在 37 文献中.
- Du X et al. ITGB4 deficiency induces mucus hypersecretion by upregulating MUC5AC in RSV-infected airway epithelial cells. Int J Biol Sci 18:349-359 (2022). PubMed: 34975337
- Du X et al. ITGB4 Deficiency in Airway Epithelium Aggravates RSV Infection and Increases HDM Sensitivity. Front Immunol 13:912095 (2022). PubMed: 35958591
- Chi L et al. N-Acetyl-L-Cysteine Protects Airway Epithelial Cells during Respiratory Syncytial Virus Infection against Mucin Synthesis, Oxidative Stress, and Inflammatory Response and Inhibits HSPA6 Expression. Anal Cell Pathol (Amst) 2022:4846336 (2022). PubMed: 36046596
- Yin B et al. Epidermal growth factor/epidermal growth factor receptor moves into the osteoblasts' cell nuclei where it is involved to regulate STAT5's signaling. Biofactors 47:351-362 (2021). PubMed: 33503297
- de Wit M et al. Mutation and drug-specific intracellular accumulation of EGFR predict clinical responses to tyrosine kinase inhibitors. EBioMedicine 56:102796 (2020). PubMed: 32512509