Anti-mTOR antibody [53E11] (ab87540)

Overview

• Product nameAnti-mTOR antibody [53E11]See all mTOR primary antibodies ...
• Description
  Mouse monoclonal [53E11] to mTOR
• Tested applicationsWB, Flow Cyt, ICC/IF more details
• Species reactivity
  Reacts with: Mouse, Rat, Human
  
• Immunogen

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

  Read Abcam's proprietary immunogen policy

• Positive controlThis 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

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: 1% BSA, PBS, pH 7.4
• Concentration information loading...
• PurityIgG fraction
• Clonality Monoclonal
• Clone number53E11
• IsotypeIgG2b
• Research Areas
  • Metabolism
  • Types of disease
  • Obesity

  • Cancer
  • Cell cycle
  • Cell cycle inhibitors
  • Other

  • Cardiovascular
  • Heart
  • Cardiogenesis
  • Transcription factors/regulators

  • Epigenetics and Nuclear Signaling
  • DNA / RNA
  • DNA Damage & Repair
  • DNA Damage Response
  • DNA Damage Recognition

  • Cell Biology
  • Cell Cycle
  • Cell Cycle Inhibitors
  • Other

Applications

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
  modificationsAutophosphorylated; 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.

Target information above from: UniProt accession P42345 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 2475 Human
  • Entrez Gene: 56717 Mouse
  • Entrez Gene: 56718 Rat
  • Omim: 601231 Human
  • SwissProt: P42345 Human
  • SwissProt: Q9JLN9 Mouse
  • SwissProt: P42346 Rat
  • Unigene: 338207 Human

  • Unigene: 21158 Mouse
  • Unigene: 11008 Rat

  see all

• Alternative names
  dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) antibodyFK506 binding protein 12 rapamycin associated protein 1 antibodyFK506 binding protein 12 rapamycin associated protein 2 antibody

  FK506 binding protein 12 rapamycin complex associated protein 1 antibodyFK506-binding protein 12-rapamycin complex-associated protein 1 antibodyFKBP rapamycin associated protein antibodyFKBP12 rapamycin complex associated protein antibodyFKBP12-rapamycin complex-associated protein antibodyFLJ44809 antibodyFRAP antibodyFRAP1 antibodyFRAP2 antibodyMammalian target of rapamycin antibodyMechanistic target of rapamycin antibodyMTOR antibodyMTOR_HUMAN antibodyOTTHUMP00000001983 antibodyRAFT1 antibodyRapamycin and FKBP12 target 1 antibodyRapamycin associated protein FRAP2 antibodyRapamycin target protein 1 antibodyRapamycin target protein antibodyRAPT1 antibodySerine/threonine-protein kinase mTOR antibody

  see all

Anti-mTOR antibody [53E11] images

• 

  Flow Cytometry-Anti-mTOR antibody [53E11](ab87540)
  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.
  
  
• 

  Western blot - mTOR antibody [53E11] (ab87540)
  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 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)
  
  Lysates/proteins at 10 µg per lane.
  
  Secondary
  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (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
• 

  Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody [53E11] (ab87540)
  ICC/IF image of ab87540 stained HepG2 cells. The cells were 4% PFA 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 (ab87540, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse 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.
• 

  Western blot - Anti-mTOR antibody [53E11] (ab87540)
  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 polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (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