Recombinant Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448)
- Suitable for: WB, IHC-P, Dot blot, IHC-Fr
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-mTOR (phospho S2448) antibody [EPR426(2)]
See all mTOR primary antibodies -
Description
Rabbit monoclonal [EPR426(2)] to mTOR (phospho S2448) -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Dot blot, IHC-Frmore details
Unsuitable for: Flow Cyt,ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat, Pig -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293, HeLa and NIH/3T3 cell lysates. IHC-P: Human breast and endometrium carcinoma tissue. Dot Blot: mTOR (phospho S2448) phospho peptide. IHC-Fr: Human heart tissue sections.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR426(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab109268 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 | (4) |
1/1000 - 1/10000. Predicted molecular weight: 289 kDa.
|
IHC-P | (1) |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
Dot blot |
1/1000.
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IHC-Fr |
Use at an assay dependent concentration.
|
Notes |
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WB
1/1000 - 1/10000. Predicted molecular weight: 289 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Dot blot
1/1000. |
IHC-Fr
Use at an assay dependent concentration. |
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 (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/2000 dilution (Purified)
Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate
Lane 3 : HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate. Then the membrane was incubated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 289 kDaBlocking buffer: 5% NFDM/TBST
Dilutiing buffer: 5% NFDM /TBST -
IHC image of mTOR (phospho S2448) staining in a section of frozen normal human heart performed on a Leica BONDTM system using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab109268, 1/200 dilution, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
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Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using unpurified ab109268 at a dilution of 1/50. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.
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Dot blot analysis of mTOR (phospho S2448) phospho peptide (Lane 1) and mTOR non-phospho peptide (Lane 2) labeling mTOR (phospho S2448) phospho peptide with purified ab109268 at a dilution of 1/1000 (0.073ug/ml). A Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) was used as the secondary antibody at a dilution of 1/100,000.
Blocking buffer: 5% NFDM/TBST
Diluting buffer: 5% NFDM /TBST -
All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)
Lane 1 : untreated NIH/3T3 cell lysate
Lane 2 : NIH/3T3 cell lysate treated with insulin
Lane 3 : NIH/3T3 cell lysate treated with insulin, then the membrane treated wth alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))
Predicted band size: 289 kDa
Observed band size: 289 kDa
Exposure time: 2 minutesBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemical cytoplasmic and nuclear staining of paraffin embedded human endometrium carcinoma with purified ab109268 at a working dilution of 1 in 100. The secondary antibody used is ab97051, a HRP goat anti-rabbit IgG (H+L), at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/1000 dilution (purified)
Lane 1 : untreated HeLa cell lysate
Lane 2 : HeLa cell lysate treated with insulin
Lane 3 : HeLa cell lystae treated with insulin, and the membrane treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (HRP goat anti-rabbit (H+L))
Predicted band size: 289 kDa
Observed band size: 289 kDa
Exposure time: 1 minuteBlocking buffer: 2% BSA/TBST
Dilution buffer: 2% BSA/TBST
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All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/2000 dilution (purified)
Lane 1 : untreated HEK293 cell lysate
Lane 2 : HEK293 cell lysate treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (HRP goat anti-rabbit IgG (H+L))
Predicted band size: 289 kDa
Observed band size: 289 kDa
Exposure time: 3 minutesBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268) at 1/5000 dilution (unpurified)
Lane 1 : untreated HEK293 cell lysate
Lane 2 : HEK293 cell lysate treated with alkaline phosphatase
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution (HRP goat anti-rabbit IgG (H+L))
Predicted band size: 289 kDa
Observed band size: 289 kDa
Exposure time: 3 minutesBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (441)
ab109268 has been referenced in 441 publications.
- Li YK et al. Validation of ESM1 Related to Ovarian Cancer and the Biological Function and Prognostic Significance. Int J Biol Sci 19:258-280 (2023). PubMed: 36594088
- 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
- Oh S et al. Non-bioenergetic roles of mitochondrial GPD2 promote tumor progression. Theranostics 13:438-457 (2023). PubMed: 36632231
- Wang X et al. The KLF4-p62 axis prevents vascular endothelial cell injury via the mTOR/S6K pathway and autophagy in diabetic kidney disease. Endokrynol Pol 73:837-845 (2022). PubMed: 36621906
- Peng J et al. The m6A methyltransferase METTL3 affects autophagy and progression of nasopharyngeal carcinoma by regulating the stability of lncRNA ZFAS1. Infect Agent Cancer 17:1 (2022). PubMed: 34980191