Overview

  • Product nameAnti-mTOR antibodySee all mTOR primary antibodies ...
  • Description
    Rabbit polyclonal to mTOR
  • Tested applicationsICC/IF, WB, IHC-Fr, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Equus
    Predicted to work with: Sheep, Rabbit, Goat, Guinea pig, Cow, Dog, Pig, Ferret, Rhesus monkey, Elephant
  • Immunogen

    Synthetic peptide (Human) - which represents a portion of human Mammalian Target of Rapamycin encoded in part by exons 5 and 6. (Peptide available as ab39393.)

Properties

Applications

Our Abpromise guarantee covers the use of ab2732 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 at an assay dependent concentration.
WB 1/2000. Can be blocked with mTOR peptide (ab39393).
IHC-Fr 1/100.
IP 1/1000.
IHC-P 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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 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

  • ab2732 at a 1:100 dilution confocally staining mTOR (red) in L6 myotubes, alongside a nuclear antigen antibody (green).    
  • mTOR protein kinase assay:

    ab2732 (1:1000 dilution) immunoprecipitates were incubated at 30°C with recombinant 4E-BP1 and 32P-gATP. Autoradiograph shows 32P incorporated into 4E-BP1.

  • ICC/IF image of ab2732 stained HepG2 cells. The cells were 100% methanol fixed (5 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 (ab2732, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • ab2732 staining rat testis sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1%BSA for 10 minutes at 21°C, followed by staining with ab2732 at 1/2000 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.  

    Positive components in the submitted image are indicated by coloured arrowheads: red for densely positive nuclei that appear to be confined to the stem cell layer (immediately above, some spermatogonia exhibit a nuclear speckling). Yellow indicates a moderate cytoplasmic positivity, more evident in some seminiferous tubules (ST). Green for a strong apparently cytoplasmic positivity at the lumenal surface of a proportion of ST.

    See Abreview

References for Anti-mTOR antibody (ab2732)

This product has been referenced in:
  • Burkhardt L  et al. CHD1 is a 5q21 tumor suppressor required for ERG rearrangement in prostate cancer. Cancer Res 73:2795-805 (2013). Read more (PubMed: 23492366) »
  • Piyanova A  et al. Loss of CB1 receptors leads to decreased cathepsin D levels and accelerated lipofuscin accumulation in the hippocampus. Mech Ageing Dev 134:391-9 (2013). Read more (PubMed: 23954857) »

See all 8 Publications for this product

Product Wall

Thank you and your customer for your inquiry.

I believe that the images that your customers provided do show very nice specific staining.

According to the Uniprot homepage (http://www.uniprot.org/uniprot/P42345) mTOR is expressed in...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Equus Tissue sections (Lung)
Specification Lung
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated
Permeabilization Yes - 0.2% Triton-X100
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 20% · Temperature: 20°C
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Abcam user community

Verified customer

Submitted May 14 2013

Thank you for contacting us.

As we deliver 50 µg at 1mg/ml, the expected volume should be 50ul.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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I am glad you have found the information I provided of help. If I can be of any further assistance, please do not hesitate to let me know.

Until then, I wish you all the best with your research.

Thank you for contacting us and your interest in our products.

When trying to detect a protein of the size of mTOR in Western blotting, there is some movement of the band that would be expected away from the predicted band size. Due to how th...

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Thank you for your email.
Your credit note ID is xxx.
I am sorry that this antibody did not perform in rat as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the in...

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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...

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Thank you for your email.
I have checked our catalog, but unfortunately we do not carry rat prostate tissue slides, only human ones.
In that case, I'd be happy to either replace the antibody (we carry several mTOR antibodies that were tested ...

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Thank you for contacting us.
The lab informed me that the human and rat sequences are the same for mTOR and thus we would expect ab2732 to work with rat tissue.
Unfortunately at this time we have not tested this antibody with rat tissue in-ho...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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