Overview

  • Product nameAnti-mTOR antibody
    See all mTOR primary antibodies
  • Description
    Rabbit polyclonal to mTOR
  • Tested applicationsICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat, Fruit fly (Drosophila melanogaster)
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 2150 - 2250 of Human mTOR.

    (Peptide available as ab45988.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A431; HEK293; HepG2 (data not shown for HepG2). This antibody gave a positive result in IHC in the following FFPE tissue: Human Testis. IF/ICC: HeLa cell line.

Properties

Applications

Our Abpromise guarantee covers the use of ab45989 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
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 250 kDa (predicted molecular weight: 289 kDa).Can be blocked with mTOR peptide (ab45988).

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 protein12-rapamycin associated protein 1 antibody
    • FK506 binding protein12-rapamycin associated protein 1 antibody
    • FK506 binding protein12-rapamycin associated protein 2 antibody
    • FK506 binding protein12-rapamycin associated protein 2 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 antibody images

  • ICC/IF image of ab45989 stained HeLa cells. The cells were 4% Formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45989, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 Goat anti-Rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of mTOR staining in Human Testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45989, 1 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-mTOR antibody (ab45989) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 Human embryonic kidney cell line Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 289 kDa
    Observed band size : 250 kDa (why is the actual band size different from the predicted?)
  • Anti-mTOR antibody (ab45989) at 1 µg/ml + Human HEK293 whole cell lysate at 12.5 µg

    Secondary
    An HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/16000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 289 kDa
    Observed band size : 289 kDa
    Additional bands at : 180 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

    Blocking Step: 5% Milk for 45 minutes at room temperature.

    See Abreview

References for Anti-mTOR antibody (ab45989)

ab45989 has not yet been referenced specifically in any publications.

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 23°C
Sample Fruit fly (Drosophila melanogaster) Cell (S2R+ cell line)
Specification S2R+ cell line
Permeabilization Yes - Igepal 0.37%
Fixative Paraformaldehyde
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Abcam user community

Verified customer

Submitted Feb 26 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 12.5 µg
Specification HEK293
Gel Running Conditions Reduced Denaturing (4-20%)
Blocking step Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: RT°C
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Abcam user community

Verified customer

Submitted Jun 08 2010

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