重组
RabMAb

Anti-ErbB 2 (phospho Y1248) + ErbB 4 (phospho Y1284)抗体[EPR19547] (ab201013)

概述

  • 产品名称
    Anti-ErbB 2 (phospho Y1248) + ErbB 4 (phospho Y1284)抗体[EPR19547]
  • 描述
    兔单克隆抗体[EPR19547] to ErbB 2 (phospho Y1248) + ErbB 4 (phospho Y1284)
  • 宿主
    Rabbit
  • 经测试应用
    适用于: WB, ICC/IF, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide within Human ErbB 2 aa 1200 to the C-terminus (phospho Y1248). The exact sequence is proprietary.
    Database link: P04626

  • 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: HeLa cells. IP: HeLa and A431 whole cell lysates starved 4 hours, then treated with 200 ng/ml EGF for 15 minutes.
  • 常规说明

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

性能

应用

Our Abpromise guarantee covers the use of ab201013 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/5000. Detects a band of approximately 180 kDa (predicted molecular weight: 138, 147 kDa).
ICC/IF 1/100.
Flow Cyt 1/50.
IP 1/30.

靶标

  • 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

图片

  • All lanes : Anti-ErbB 2 (phospho Y1248) + ErbB 4 (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).

  • All lanes : Anti-ErbB 2 (phospho Y1248) + ErbB 4 (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 seconds


    Blocking/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)

  • 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.

  • 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 Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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.

文献

ab201013 has not yet been referenced specifically in any publications.

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