Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-mTOR antibody [EPR390(N)] (ab134903)

Knockout Tested Rabbit recombinant monoclonal mTOR antibody [EPR390(N)]. Validated in WB and tested in Mouse, Rat, Human. Cited in 7 publication(s).

Overview

  • Product name

    Anti-mTOR antibody [EPR390(N)]
    See all mTOR primary antibodies
  • Description

    Rabbit monoclonal [EPR390(N)] to mTOR
  • Host species

    Rabbit
  • Specificity

    Expression levels of the target protein vary with sample type and some optimisation may be required.

  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF,IHC-P or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues near the N terminus of Human mTOR (UniProt ID: P42345).

  • Positive control

    • WB: Human brain, heart, liver and kidney lysates, Mouse brain heart, liver and kidney lysates, Rat brain, heart and kidney lysates, HeLa, MCF7, K562, Raji, HepG2, and 293T cell lysates
  • General notes

     

     

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab134903 in the following tested applications.

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

Application Abreviews Notes
WB 1/10000. Predicted molecular weight: 289 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF,IHC-P or IP.
  • Target

    • Function

      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'.
    • Tissue specificity

      Expressed in numerous tissues, with highest levels in testis.
    • Sequence similarities

      Belongs to the PI3/PI4-kinase family.
      Contains 1 FAT domain.
      Contains 1 FATC domain.
      Contains 7 HEAT repeats.
      Contains 1 PI3K/PI4K domain.
    • Post-translational
      modifications

      Autophosphorylated; when part of mTORC1 or mTORC2.
    • Cellular localization

      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
    • Database links

    • Alternative names

      • 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

    Images

    • All lanes : Anti-mTOR antibody [EPR390(N)] (ab134903) at 1/10000 dilution

      Lane 1 : Wild-type HEK293T whole cell lysate
      Lane 2 : MTOR knockout HEK293T whole cell lysate
      Lane 3 : K562 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 289 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab134903 observed at 289 kDa. Red - loading control, ab130007, observed at 130 kDa.

      ab134903 was shown to specifically react with mTOR in wild-type HEK293T cells as signal was lost in MTOR knockout cells. Wild-type and MTOR knockout samples were subjected to SDS-PAGE. Ab134903 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-mTOR antibody [EPR390(N)] (ab134903) at 1/1000 dilution

      Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
      Lane 3 : Human brain lysates
      Lane 4 : Mouse brain lysates
      Lane 5 : Rat brain lysates
      Lane 6 : Human liver lysates
      Lane 7 : Mouse liver lysates
      Lane 8 : Human heart lysates
      Lane 9 : Mouse heart lysates
      Lane 10 : Rat heart lysates
      Lane 11 : Human kidney lysates
      Lane 12 : Mouse kidney lysates
      Lane 13 : Rat kidney lysates

      Lysates/proteins at 20 µg per lane.

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

      Predicted band size: 289 kDa
      Observed band size: 290 kDa
      why is the actual band size different from the predicted?



      Blocking buffer and concentration 5% NFDM/TBST
      Diluting buffer and concentration 5% NFDM/TBST

      Exposure time:
      Lane 1 to 5: 80 seconds
      Lane 6 to 13: 180 seconds

      This antibody detects non-specific bands. It doesn’t detect the target band in some mouse and rat tissues.

    • All lanes : Anti-mTOR antibody [EPR390(N)] (ab134903) at 1/10000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : K562 cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : HepG2 cell lysate
      Lane 5 : 293T cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit polyclonal IgG at 1/2000 dilution

      Predicted band size: 289 kDa
      Observed band size: 289 kDa

    References

    This product has been referenced in:

    • Adimonye A  et al. Analysis of the PI3K-AKT-mTOR pathway in penile cancer: evaluation of a therapeutically targetable pathway. Oncotarget 9:16074-16086 (2018). Read more (PubMed: 29662627) »
    • Ma D  et al. Ketogenic diet enhances neurovascular function with altered gut microbiome in young healthy mice. Sci Rep 8:6670 (2018). Read more (PubMed: 29703936) »
    See all 11 Publications for this product

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