Recombinant
RabMAb

Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Overview

  • Product name
    Anti-mTOR (phospho S2448) antibody [EPR426(2)]
    See all mTOR primary antibodies
  • Description
    Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448)
  • Specificity
    This antibody only detects mTOR/FRAP phosphorylated at serine 2448.
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity
    Reacts with: Mouse, Human, Pig
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human mTOR (phospho S2448). The exact sequence is proprietary.

  • Positive control
    • WB: 293T, HeLa and NIH/3T3 cell lysates. IHC-P: Human breast carcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Alternative versions available:

    Anti-mTOR (phospho S2448) antibody (Alexa Fluor® 488) [EPR426(2)] (ab197237)
    Anti-mTOR (phospho S2448) antibody (Alexa Fluor® 647) [EPR426(2)] (ab196913)
    Anti-mTOR (phospho S2448) antibody (HRP) [EPR426(2)] (ab196914)

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

Properties

Applications

Our Abpromise guarantee covers the use of ab109268 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 1/1000 - 1/10000. Predicted molecular weight: 289 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF 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

    • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab109268 at a dilution of 1/50.

    • All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)

      Lane 1 : untreated NIH/3T3 cell lysate
      Lane 2 : NIH/3T3 cell lysate treated with insulin
      Lane 3 : NIH/3T3 cell lysate treated with insulin, then the membrane treated wth alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

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

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


      Exposure time : 2 minutes

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • Immunohistochemical cytoplasmic and nuclear staining of paraffin embedded human endometrium carcinoma with purified ab109268 at a working dilution of 1 in 100. The secondary antibody used is ab97051, a HRP goat anti-rabbit IgG (H+L), at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

    • All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)

      Lane 1 : untreated HeLa cell lysate
      Lane 2 : HeLa cell lysate treated with insulin
      Lane 3 : HeLa cell lystae treated with insulin, and the membrane treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (HRP goat anti-rabbit (H+L))

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


      Exposure time : 1 minute

      Blocking buffer: 2% BSA/TBST

      Dilution buffer: 2% BSA/TBST

    • All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/2000 dilution (purified)

      Lane 1 : untreated HEK293 cell lysate
      Lane 2 : HEK293 cell lysate treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

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

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


      Exposure time : 3 minutes

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/5000 dilution (unpurified)

      Lane 1 : untreated HEK293 cell lysate
      Lane 2 : HEK293 cell lysate treated with alkaline phosphatase

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (HRP goat anti-rabbit IgG (H+L))

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


      Exposure time : 3 minutes

      Blocking buffer: 5% NFDM/TBST

      Dilution buffer: 5% NFDM/TBST

    References

    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) »
    • Huang G  et al. Notoginsenoside R1 attenuates glucose-induced podocyte injury via the inhibition of apoptosis and the activation of autophagy through the PI3K/Akt/mTOR signaling pathway. Int J Mol Med 39:559-568 (2017). Human . Read more (PubMed: 28112381) »

    See all 33 Publications for this product

    Customer reviews and Q&As

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: EDTA, pH8, 20 min 100C
    Sample
    Human Tissue sections (breast cancer)
    Specification
    breast cancer
    Permeabilization
    No
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Feb 26 2015

    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing (4-12)
    Sample
    Mouse Cell lysate - whole cell (placenta)
    Specification
    placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Varkha Agrawal

    Verified customer

    Submitted Jul 14 2014

    Tween-20 is often used at 0.2%-0.5% as a membrane permeabilizing solution. At 0.1% in PBS, Tween-20 can help reduce background but does not have much effect for permeabilizing. Tween-20 may be found at many life science vendors.

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

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