RabMAb

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

Overview

  • Product nameAnti-mTOR (phospho S2448) antibody [EPR426(2)]See all mTOR primary antibodies ...
  • Description
    Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448)
  • SpecificityThis antibody only detects mTOR/FRAP phosphorylated at serine 2448.
  • Tested applicationsWB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Phospho specific peptide corresponding to residues surrounding Serine 2448 of Human mTOR.

  • Positive control
    • 293T cell lysate; Human breast carcinoma tissue; HeLa cells.
  • General notesProduced under U.S. Patent No. 5,675,063.

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.
ICC/IF 1/100 - 1/250.
Flow Cyt 1/100.
  • Application notesIs unsuitable for IP.
  • Target

    • FunctionKinase 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 specificityExpressed in numerous tissues, with highest levels in testis.
    • Sequence similaritiesBelongs 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 localizationEndoplasmic 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 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

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

    • Overlay histogram showing HeLa cells stained with ab109268 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab109268, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution

      Lane 1 : 293T cell lysate, untreated
      Lane 2 : 293T cell lysate, treated with Alkaline Phosphatase

      Lysates/proteins at 10 µg per lane.


      Predicted band size : 289 kDa
    • Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue using ab109268 at a dilution of 1/50.
    • Immunofluorescent staining of HeLa cells using ab109268 at a dilution of 1/100.

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

    This product has been referenced in:
    • Duan Y  et al. Effects of tonic immobility (TI) and corticosterone (CORT) on energy status and protein metabolism in pectoralis major muscle of broiler chickens. Comp Biochem Physiol A Mol Integr Physiol 169:90-5 (2014). Chicken . Read more (PubMed: 24389091) »
    • Vallejo D  et al. Autophagic response in the Rabbit Hemorrhagic Disease, an animal model of virally-induced fulminant hepatic failure. Vet Res 45:15 (2014). WB ; Rabbit . Read more (PubMed: 24490870) »

    See all 7 Publications for this product

    Product Wall

    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"