Overview

  • Product name
    Anti-mTOR antibody [53E11]
    See all mTOR primary antibodies
  • Description
    Mouse monoclonal [53E11] to mTOR
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment derived from within residues 1 - 350 of Human mTOR.

  • Positive control
    • This antibody gave a positive signal in the following lysates: HeLa whole cell; Jurkat whole cell; MCF7 whole cell; Hek293 whole cell; A431 whole cell; PC12 whole cell; Human liver tissue; Mouse testis; Rat testis. Flow Cytometry: HeLa cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    Alternative versions available:

    Anti-mTOR antibody [53E11] - BSA and Azide free (ab171829)
    Anti-mTOR antibody (HRP) [53E11] (ab203646)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab87540 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 Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 265 kDa (predicted molecular weight: 289 kDa).
Flow Cyt Use 1µg for 106 cells.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration. PubMed: 21900751

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 [53E11] (ab87540) at 5 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 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 5 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 6 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 289 kDa
    Observed band size : 265 kDa (why is the actual band size different from the predicted?)


    Exposure time : 4 minutes
  • Overlay histogram showing HeLa cells stained with ab87540 (red line). The cells were fixed with 80% methanol (5 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 (ab87540, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2b [PLPV219] (ab91366, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • All lanes : Anti-mTOR antibody [53E11] (ab87540) at 1 µg/ml

    Lane 1 : Testis (Mouse) Tissue Lysate
    Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 3 : Testis (Rat) Tissue Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 289 kDa
    Observed band size : 265 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 31 kDa,50 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes

References

This product has been referenced in:
  • Imasawa T  et al. High glucose repatterns human podocyte energy metabolism during differentiation and diabetic nephropathy. FASEB J 31:294-307 (2017). WB ; Human . Read more (PubMed: 27825100) »
  • Cui N  et al. Role of Triggering Receptor Expressed on Myeloid Cell-1 Expression in Mammalian Target of Rapamycin Modulation of CD8(+) T-cell Differentiation during the Immune Response to Invasive Pulmonary Aspergillosis. Chin Med J (Engl) 130:1211-1217 (2017). Read more (PubMed: 28485322) »

See all 3 Publications for this product

Customer reviews and Q&As

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Muscle)
Gel Running Conditions
Reduced Denaturing (8%)
Loading amount
30 µg
Specification
Muscle
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jan 04 2016

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Sample
Human Cell (muscle)
Specification
muscle
Permeabilization
Yes - Triton
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Nov 18 2014

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...

Read More

Thank you for your inquiry.

Please assure your customer that we do guarantee that this antibody works in ICC/IF with our Abpromise.

We did not test ab87540 ourselves in ICC/IF but added it to the applications list after it wasused i...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (human fibroblast)
Loading amount
50 µg
Specification
human fibroblast
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 14 2011

Thank you for confirming the order 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...

Read More

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

Sign up