Name: Anti-mTOR (phospho S2448) antibody [EPR426(2)]
Brand: RabMAb
SKU: ab109268
Rating: 4 (5 reviews)

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Western blot - 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

Carrier Free HRP

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: Pig
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR426(2)
Isotype

IgG
Research areas
- Neuroscience
- Development
- Neuroscience
- Processes

Alternative Versions
- Anti-mTOR (phospho S2448) antibody [EPR426(2)] - BSA and Azide free (ab177734)
- HRP Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab196914)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human mTOR protein (ab114179)

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	(3)	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.
IHC-Fr		Use at an assay dependent concentration.

Notes
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.

Application notes

Is unsuitable for Flow Cyt,ICC/IF or IP.

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.
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
- Omim: 601231 Human
- SwissProt: P42345 Human
- SwissProt: Q9JLN9 Mouse
- Unigene: 338207 Human
- Unigene: 21158 Mouse
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

Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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 kDa

Blocking buffer: 5% NFDM/TBST
Dilutiing buffer: 5% NFDM /TBST
Immunohistochemistry (Frozen sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

IHC image of mTOR (phospho S2448) staining in a section of frozen normal human heart performed on a Leica BOND^TM 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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.
Dot Blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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.
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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 minute

Blocking buffer: 2% BSA/TBST
Dilution buffer: 2% BSA/TBST
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

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 minutes

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

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Datasheet download

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Publishing research using ab109268? Please let us know so that we can cite the reference in this datasheet.

ab109268 has been referenced in 334 publications.

Xie B et al. Induction of autophagy and suppression of type I IFN secretion by CSFV. Autophagy 17:925-947 (2021). PubMed: 32160078
Hu Y et al. Long noncoding RNA TMPO-AS1/miR-126-5p/BRCC3 axis accelerates gastric cancer progression and angiogenesis via activating PI3K/Akt/mTOR pathway. J Gastroenterol Hepatol 36:1877-1888 (2021). PubMed: 33295056
Xu J et al. Ubiquitously specific protease 4 inhibitor-Vialinin A attenuates inflammation and fibrosis in S100-induced hepatitis mice through Rheb/mTOR signalling. J Cell Mol Med 25:1140-1150 (2021). PubMed: 33295107
Cho J et al. Vascular remodeling by placenta-derived mesenchymal stem cells restores ovarian function in ovariectomized rat model via the VEGF pathway. Lab Invest 101:304-317 (2021). PubMed: 33303971
Qiu X et al. Metformin alleviates ß-glycerophosphate-induced calcification of vascular smooth muscle cells via AMPK/mTOR-activated autophagy. Exp Ther Med 21:58 (2021). PubMed: 33365058

View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application

Western blot

Sample

Mouse Cell lysate - whole cell (primary cultured mouse cortical neurons)

Gel Running Conditions

Reduced Denaturing

Loading amount

200000 cells

Specification

primary cultured mouse cortical neurons

Blocking step

BSA as blocking agent for 45 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 02 2020

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (DU145 Prostate cancer cell line)

Permeabilization

Yes - 0.1% Triton X-100

Specification

DU145 Prostate cancer cell line

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Fixative

Paraformaldehyde

DR. Dimitra Kalamida

Verified customer

Submitted Dec 28 2019

Application

Western blot

Sample

Human Cell lysate - whole cell (C42B Prostate cancer cell line)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

C42B Prostate cancer cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

DR. Dimitra Kalamida

Verified customer

Submitted Dec 25 2019

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

H2O2 as blocking agent for 10 minute(s) · Concentration: 3% · Temperature: 25°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: EDTA, pH8, 20 min 100C

Sample

Human Tissue sections (breast cancer)

Specification

breast cancer

Permeabilization

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Feb 26 2015

Application

Western blot

Loading amount

20 µg

Gel Running Conditions

Reduced Denaturing (4-12)

Sample

Mouse Cell lysate - whole cell (placenta)

Specification

placenta

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Varkha Agrawal

Verified customer

Submitted Jul 14 2014

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (334)

Customer reviews and Q&As

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Western blot abreview for Anti-mTOR (phospho S2448) antibody [EPR426(2)]

Immunocytochemistry/ Immunofluorescence abreview for Anti-mTOR (phospho S2448) antibody [EPR426(2)]

Western blot abreview for Anti-mTOR (phospho S2448) antibody [EPR426(2)]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-mTOR (phospho S2448) antibody [EPR426(2)]

Western blot abreview for Anti-mTOR (phospho S2448) [EPR426(2)] antibody

Post-translational
modifications