リコンビナント
RabMAb

Anti-mTOR 抗体 [Y391] (ab32028)

製品の概要

  • 製品名
    Anti-mTOR antibody [Y391]
    mTOR 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [Y391] to mTOR
  • アプリケーション
    適用あり: ChIP, WB, IP, IHC-Pmore details
    適用なし: Flow Cyt or ICC/IF
  • 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 mTOR aa 2400-2500 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab193663)

  • エピトープ
    ab32028 reacts with an epitope located in the C terminal region of mTOR.
  • ポジティブ・コントロール
    • WB: HeLa, Jurkat and Raw264.7 cell lysates and rat brain tissue lysate. IHC-P: Human breast carcinoma, mouse testis and rat testis tissues. IP: Rat brain tissue lysate and HeLa cell lysate.
  • 特記事項

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32028 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
ChIP Use at an assay dependent concentration. PubMed: 20233713
WB 1/1000 - 1/5000. Detects a band of approximately 250 kDa (predicted molecular weight: 289 kDa).Can be blocked with mTOR peptide (ab193663).
IP 1/50 - 1/100.
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

  • 追加情報
    Is unsuitable for Flow Cyt or ICC/IF.
  • ターゲット情報

    • 機能
      Kinase subunit of both mTORC1 and mTORC2, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-421', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Phosphorylates MAF1 leading to attenuation of its RNA polymerase III-repressive function. mTORC2 is also activated by growth. factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. mTORC2 plays a critical role in AKT1 'Ser-473' phosphorylation, which may facilitate the phosphorylation of the activation loop of AKT1 on 'Thr-308' by PDK1 which is a prerequisite for full activation. mTORC2 regulates the phosphorylation of SGK1 at 'Ser-422'. mTORC2 also modulates the phosphorylation of PRKCA on 'Ser-657'.
    • 組織特異性
      Expressed in numerous tissues, with highest levels in testis.
    • 配列類似性
      Belongs to the PI3/PI4-kinase family.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 7 HEAT repeats.
      Contains 1 PI3K/PI4K domain.
    • 翻訳後修飾
      Autophosphorylated; when part of mTORC1 or mTORC2.
    • 細胞内局在
      Endoplasmic reticulum membrane. Golgi apparatus membrane. Mitochondrion outer membrane. Lysosome. Cytoplasm. Nucleus > PML body. Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB.
    • Information by UniProt
    • 参照データベース
    • 別名
      • dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) antibody
      • FK506 binding protein 12 rapamycin associated protein 1 antibody
      • FK506 binding protein 12 rapamycin associated protein 2 antibody
      • FK506 binding protein 12 rapamycin complex associated protein 1 antibody
      • FK506-binding protein 12-rapamycin complex-associated protein 1 antibody
      • FKBP rapamycin associated protein antibody
      • FKBP12 rapamycin complex associated protein antibody
      • FKBP12-rapamycin complex-associated protein 1 antibody
      • FKBP12-rapamycin complex-associated protein antibody
      • FLJ44809 antibody
      • FRAP antibody
      • FRAP1 antibody
      • FRAP2 antibody
      • Mammalian target of rapamycin antibody
      • Mechanistic target of rapamycin antibody
      • mTOR antibody
      • MTOR_HUMAN antibody
      • OTTHUMP00000001983 antibody
      • RAFT1 antibody
      • Rapamycin and FKBP12 target 1 antibody
      • Rapamycin associated protein FRAP2 antibody
      • Rapamycin target protein 1 antibody
      • Rapamycin target protein antibody
      • RAPT1 antibody
      • Serine/threonine-protein kinase mTOR antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • All lanes : Anti-mTOR antibody [Y391] (ab32028) at 1/5000 dilution (purified)

      Lane 1 : HeLa whole cell lysate
      Lane 2 : Jurkat whole cell lysate
      Lane 3 : Rat brain tissue lysate
      Lane 4 : Raw264.7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 289 kDa
      Observed band size : 289 kDa
      Blocking and dilution buffer: 5% NFDM/TBST
    • ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in HeLa whole cell lysate.

      Lane 1 (input): HeLa whole cell lysate (10µg)

      Lane 2 (+): ab32028 + HeLa whole cell lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32028 in HeLa whole cell lysate.

      For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    • ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in rat brain tissue lysate.

      Lane 1 (input): Rat brain tissue lysate (10µg)

      Lane 2 (+): ab32028 + rat brain tissue lysate.

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32028 in rat brain tissue lysate.

      For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    参考文献

    This product has been referenced in:
    • Zhu H  et al. IDH1 R132H Mutation Enhances Cell Migration by Activating AKT-mTOR Signaling Pathway, but Sensitizes Cells to 5-FU Treatment as NADPH and GSH Are Reduced. PLoS One 12:e0169038 (2017). WB ; Human . Read more (PubMed: 28052098) »
    • Lin Q  et al. Cisplatin regulates cell autophagy in endometrial cancer cells via the PI3K/AKT/mTOR signalling pathway. Oncol Lett 13:3567-3571 (2017). Read more (PubMed: 28521459) »

    See all 23 Publications for this product

    レビューと Q&A

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (prostate cancer)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA, pH9, 100C 20 min
    Permeabilization
    No
    Specification
    prostate cancer
    Blocking step
    H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    投稿 Apr 24 2015

    Thank you for your reply.

    As requested, I am sending you a new vial of mTOR antibody, ab2732 to use in your IP experiments, since the original vial of antibody was not working as well as it should have. Your new order number, for tracking purp...

    Read More

    Thank you for your patience whilst I was looking in to this matter.

    I checked the concentration for this lot and it is around 1.758 mg/ml. So, I don't believe it is a concentration issue.

    However, I did notice that IP testing was onl...

    Read More

    Thank you for contacting Abcam.

    I am sorry that you are having issues with ab32028 in IP. The antibody is covered under our Abpromise for six months and is guaranteed to work in IP on mouse, rat and human samples . If we cannot resolve the iss...

    Read More

    Thank you for your email.
    I have heard back regarding ab32028:
    The lab tested WB on C6 and PC-12 cell lysates as well as on rat brain tissue lysates. They did not test rat tissues in IHC-P.
    As soon as I have more information for the othe...

    Read More

    Thank you for your reply.
    The following mTOR antibodies have been tested in rat as well as IHC-P:
    ab32028, ab51089, ab59299, and ab59350.
    The antibody ab32028 has been tested in the lab on IHC-P. I am not certain that rat tissue was used...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HUVEC)
    Loading amount
    45 µg
    Specification
    HUVEC
    Treatment
    FGF2 50 ng/ml for 5 minutes
    Gel Running Conditions
    Reduced Denaturing (gradient 4-15%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    投稿 Sep 17 2008

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (ventricular tissue lysates)
    Loading amount
    100 µg
    Specification
    ventricular tissue lysates
    Gel Running Conditions
    Reduced Denaturing (4 ~ 12% gradient NuPAGE)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
    Username

    Abcam user community

    Verified customer

    投稿 Jul 05 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Keratinocyte)
    Loading amount
    15 µg
    Specification
    Keratinocyte
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Username

    Prof. Paul Townsend

    Verified customer

    投稿 Jun 26 2008

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

    登録