Anti-mTOR antibody (ab2732)
Key features and details
- Rabbit polyclonal to mTOR
- Suitable for: ICC/IF, IP, WB
- Reacts with: Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-mTOR antibody
See all mTOR primary antibodies -
Description
Rabbit polyclonal to mTOR -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IP, WBmore details -
Species reactivity
Reacts with: Rat, Human
Predicted to work with: Mouse -
Immunogen
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Positive control
- ICC/IF: HepG2 cells. L6 myotubes. IP: HeLa lysates. WB: HeLa, HEK-293T, Jurkat-HepG2 and LNCaP whole cell lysate.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7
Preservative: 0.1% Sodium azide
Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris -
Concentration information loading...
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Purification notes
Affinity purified using the immunising peptideimmobilized on solid support. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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ICC/IF | (1) |
1/100.
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IP |
Use at 2-10 µg/mg of lysate.
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WB | (7) |
1/2000 - 1/10000. Predicted molecular weight: 289 kDa.
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Notes |
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ICC/IF
1/100. |
IP
Use at 2-10 µg/mg of lysate. |
WB
1/2000 - 1/10000. Predicted molecular weight: 289 kDa. |
Target
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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
modificationsAutophosphorylated; 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
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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
see all -
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
see all
Images
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All lanes : Anti-mTOR antibody (ab2732) at 1/2000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : MTOR [homo] CRISPR-Cas9 edited A549 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : HEK-293 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 289 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-mTOR antibody staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab2732 was shown to bind specifically to mTOR. A band was observed at 250 kDa in wild-type A549 cell lysates with no signal observed at this size in MTOR CRISPR-Cas9 edited cell line ab283257. The band observed in the CRISPR-Cas9 edited lysate lane below 250 kDa is likely to represent a truncated form of mTOR. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and MTOR CRISPR-Cas9 edited A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-mTOR antibody (ab2732) at 0.1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 5 : LNCaP (Human prostate cancer cell line) whole cell lysate
Lysates/proteins at 50 µg per lane.
Predicted band size: 289 kDa
Exposure time: 3 minutesLysates prepared using NETN lysis buffer.
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mTOR was immunoprecipitated from HeLa cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab2732 at 3 µg per reaction. mTOR was also immunoprecipitated by ab2833. Western blot was performed from the immunoprecipitate with ab2732 at 1 µg/ml.
Lane 1: ab2833 IP in HeLa whole cell lysate.
Lane 2: ab2732 IP in HeLa whole cell lysate.
Lane 3: Control IgG IP in HeLa whole cell lysate.Detection: Chemiluminescence
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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.
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ab2732 at a 1:100 dilution confocally staining mTOR (red) in L6 myotubes, alongside a nuclear antigen antibody (green).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (331)
ab2732 has been referenced in 331 publications.
- Zhu F et al. Study on the treatment of postmenopausal osteoporosis with quercetin in Liuwei Dihuang Pill based on network pharmacology. J Orthop Surg Res 18:21 (2023). PubMed: 36624462
- Xie Z et al. C2orf40 inhibits metastasis and regulates chemo-resistance and radio-resistance of nasopharyngeal carcinoma cells by influencing cell cycle and activating the PI3K/AKT/mTOR signaling pathway. J Transl Med 20:264 (2022). PubMed: 35676661
- Liu M et al. USP18 contributes to the proliferation and migration of ovarian cancer cells by regulating the AKT/mTOR signaling pathway. Acta Biochim Pol 69:417-422 (2022). PubMed: 35696596
- Abdelkader NF et al. Empagliflozin mitigates type 2 diabetes-associated peripheral neuropathy: a glucose-independent effect through AMPK signaling. Arch Pharm Res 45:475-493 (2022). PubMed: 35767208
- Grüninger PK et al. Functional characterization of the PI3K/AKT/MTOR signaling pathway for targeted therapy in B-precursor acute lymphoblastic leukemia. Cancer Gene Ther 29:1751-1760 (2022). PubMed: 35794338