Anti-ERK1 (phospho T202) + ERK2 (phospho T185) 抗体 [EPR19401] (ab201015)

製品の概要

  • 製品名Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401]
  • 製品の詳細
    Rabbit monoclonal [EPR19401] to ERK1 (phospho T202) + ERK2 (phospho T185)
  • アプリケーション適用あり: ICC/IF, IP, IHC-P, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ERK2 aa 150-250 (phospho T185). The exact sequence is proprietary.
    Database link: P28482

  • ポジティブ・コントロール
    • WB: Jurkat, treated with 200 ng/ml PMA for 30 minutes whole cell lysate; NIH/3T3, untreated and treated with 50 ng/ml PDGF for 40 minutes whole cell lysates; PC-12, untreated and treated with 200 ng/ml NGF for 4 days whole cell lysates. IHC-P: Human breast and glioma tissues. IP: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate; PC-12 treated with 100ng/ml NGF for 10min whole cell lysate. ICC/IF:Jurkat
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab201015 in the following tested applications.

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

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent concentration.
IP 1/30.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

IHC is recommended for human only.

WB 1/1000. Detects a band of approximately 44, 42 kDa (predicted molecular weight: 41 kDa).

ターゲット情報

Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] 画像

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling ERK1 (phospho T202) and ERK2 (phospho T185)

    ERK1 (phospho T202) + ERK2 (phospho T185) with ab201015 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing staining on M phase cells (PMID:26529125). After PMA treatment (200 ng/ml, 30min), the staining was increased, and LP treatment decreased the PMA induced staining.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab201015 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear with weak cytoplasm staining on Human glioma is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution

    Lane 1 : Untreated Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) treated with 200 ng/ml PMA for 30 minutes whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 41 kDa
    Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) at 1/1000 dilution

    Lane 1 : Untreated NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50 ng/ml PDGF for 40 minutes whole cell lysate
    Lane 3 : Untreated PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 200 ng/ml NGF for 4 days whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 41 kDa
    Observed band size : 42,44 kDa (why is the actual band size different from the predicted?)


    Exposure time : 10 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The induction conditions refer to PMID:12454035; PMID:17026715; PMID:22206868.

  • Dot blot analysis of ERK2 (phospho T185) labeled with ab201015 at 1/1000 dilution.

    Lane 1: ERK2 (pT185) phospho peptide: DHTGFLT(p)EYVATR  aa179-191 peptide;

    Lane 2: ERK2 Non-phospho peptide: DHTGFLTEYVATR  aa179-191 peptide;

    Lane 3: ERK2 (pY187) phospho peptide: DHTGFLTEY(p)VATR  aa179-191 peptide.

    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051)  at 1/100000 dilution was used as secondary antibody.

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Immunohistochemical analysis of paraffin-embedded hHman breast tissue labeling ERK2 (phospho T185) with ab201015 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 50ng/ml PDGF for 40min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

    Lane 1: NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate, 10µg (Input).

    Lane 2: ab201015 IP in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in NIH/3T3 treated with 50ng/ml PDGF for 40min whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • ERK2 (phospho T185) was immunoprecipitated from 0.35 mg of PC-12 (Rat adrenal gland pheochromocytoma cell line) treated with 100ng/ml NGF for 10min whole cell lysate with ab201015 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab201015 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

    Lane 1: PC-12 treated with 100ng/ml NGF for 10min whole cell lysate, 10µg (Input).

    Lane 2: ab201015 IP in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab201015 in PC-12 treated with 100ng/ml NGF for 10min whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

Anti-ERK1 (phospho T202) + ERK2 (phospho T185) antibody [EPR19401] (ab201015) 使用論文

This product has been referenced in:
  • Ferraiuolo RM  et al. The cyclin-like protein, SPY1, regulates the ERa and ERK1/2 pathways promoting tamoxifen resistance. Oncotarget 8:23337-23352 (2017). WB ; Human . Read more (PubMed: 28423577) »
  • Zhai W  et al. A1 adenosine receptor attenuates intracerebral hemorrhage-induced secondary brain injury in rats by activating the P38-MAPKAP2-Hsp27 pathway. Mol Brain 9:66 (2016). WB ; Rat . Read more (PubMed: 27301321) »

See all 2 Publications for this product

Product Wall

There are currently no Abreviews or Questions for ab201015.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"