重组Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284)抗体[EPR19547] (ab201013)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19547] to ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284)
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra), Dot blot
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284)抗体[EPR19547]
参阅全部 ErbB 2+ErbB 4 一抗 -
描述
兔单克隆抗体[EPR19547] to ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IF, IP, Flow Cyt (Intra), Dot blotmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HeLa, SK-BR-3 and A431 whole cell lysates starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. ICC/IF: HeLa cells treated with EGF (200ng/ml; 30min). Flow Cyt (intra): HeLa cells. IP: HeLa and A431 whole cell lysates starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes. Dot blot: ErbB4 non-phospho peptide, ErbB4 Y1284 phospho peptide, ErbB2 non-phospho peptide, ErbB2 Y1248 phospho peptide
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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.
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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR19547 -
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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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab201013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 - 1/5000. Detects a band of approximately 180 kDa (predicted molecular weight: 138, 147 kDa).
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ICC/IF |
1/100.
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IP |
1/30.
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Flow Cyt (Intra) |
1/50.
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Dot blot |
Use at an assay dependent concentration.
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说明 |
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WB
1/1000 - 1/5000. Detects a band of approximately 180 kDa (predicted molecular weight: 138, 147 kDa). |
ICC/IF
1/100. |
IP
1/30. |
Flow Cyt (Intra)
1/50. |
Dot blot
Use at an assay dependent concentration. |
靶标
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细胞定位
ErbB 2: Cytoplasm. Nucleus and Cell membrane. Cytoplasm, perinuclear region. Nucleus. Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1. ErbB 4: Membrane and Nucleus. Following proteolytical processing E4ICD (E4ICD1 or E4ICD2 generated from the respective isoforms) is translocated to the nucleus. Significantly more E4ICD2 than E4ICD1 is found in the nucleus. E4ICD2 colocalizes with YAP1 in the nucleus. -
数据库链接
- Entrez Gene: 2064 Human
- Entrez Gene: 2066 Human
- Omim: 164870 Human
- Omim: 600543 Human
- SwissProt: P04626 Human
- SwissProt: Q15303 Human
- Unigene: 390729 Human
- Unigene: 446352 Human
图片
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Dot blot analysis using 1/1000 dilution ab201013 and Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary at 1/100000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST
Lane 1: ErbB4 non-phospho peptide
Lane 2: ErbB4 Y1284 phospho peptide
Lane 3: ErbB2 non-phospho peptide
Lane 4: ErbB2 Y1248 phospho peptide
Exposure time: 3 minutes
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All lanes : Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] (ab201013) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lane 3 : Untreated A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 138, 147 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 and 2: 5 seconds; Lane 3 and 4: 10 seconds.
This target could be induced to increase by EGF as proved by literatures (PMID: 18945363; PMID: 17030621).
The MW is also consistent with some literatures (PMID: 18180459).
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All lanes : Anti-ErbB2 / HER2 (phospho Y1248) + ErbB4 / HER4 (phospho Y1284) antibody [EPR19547] (ab201013) at 1/5000 dilution
Lane 1 : Untreated SK-BR-3 (human mammary gland adenocarcinoma ) whole cell lysate
Lane 2 : SK-BR-3(human mammary gland adenocarcinoma ) whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 138, 147 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Diluting buffer 5% NFDM/TBST.
This target could be induced to increase by EGF that had be proved by literatures(PMID: 18945363; PMID: 17030621).
The MW is also consist with some literatures showed(PMID: 18180459)
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ERBB2 (phospho Y1248) + ERBB4 (phospho Y1284) with ab201013 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing increased cytoplasmic staining after EGF treatment (200ng/ml; 30min).
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab201013 at 1/100 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ERBB2 (phospho Y1248) + ERBB4 (phospho Y1284) with ab201013 at 1/50 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A]-Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluorr® 488) at 1/2000 dilution was used as the secondary antibody.
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ERBB2 (phospho Y1248) + ERBB4 (phospho Y1284) were immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes, with ab201013 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab201013 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: HeLa whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes 10µg (Input).
Lane 2: ab201013 IP in HeLa whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes.
Lane 3: Rabbit IgG,monoclonal [EPR25A]- Isotype Control (ab172730) instead of ab201013 in HeLa whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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ERBB2 (phospho Y1248) + ERBB4 (phospho Y1284) were immunoprecipitated from 0.35 mg of A431 (Human epidermoid carcinoma cell line) whole cell lysate, starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes, with ab201013 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab201013 at 1/500 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes 10µg (Input).
Lane 2: ab201013 IP in A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab201013 in A431 whole cell lysate starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (0)
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