Name: Anti-mTOR antibody [Y391]
SKU: ab32028
Rating: 3 (4 reviews)

Anti-mTOR antibody [Y391] (ab32028) is a rabbit monoclonal antibody detecting mTOR in Western Blot, IP, IHC-P, IHC-Fr. Suitable for Human, Mouse, Rat.

- Biophysical QC for unrivalled batch-batch consistency

- Over 230 publications

- Trusted since 2006

Images

Western blot - Anti-mTOR antibody [Y391] (AB32028), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	Flow Cyt	WB	ICC/IF	IHC-Fr
Human	Tested	Tested	Not recommended	Tested	Not recommended	Tested
Mouse	Tested	Expected	Not recommended	Tested	Not recommended	Expected
Rat	Tested	Tested	Not recommended	Tested	Not recommended	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/400	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Rat	Dilution info 1/400	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.
Species Human	Dilution info 1/400	Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/50 - 1/100	Notes -
Species Human	Dilution info 1/50 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Rat	Dilution info -	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Species Human	Dilution info -	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/5000	Notes This antibody detects non-specific bands and high background. It doesn't detect the target band in some mouse and rat tissues.
Species Rat	Dilution info 1/1000 - 1/5000	Notes This antibody detects non-specific bands and high background. It doesn't detect the target band in some mouse and rat tissues.
Species Human	Dilution info 1/1000 - 1/5000	Notes This antibody detects non-specific bands and high background. It doesn't detect the target band in some mouse and rat tissues.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/200	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information MTOR

Function

The protein expressed by the MTOR gene is a serine/threonine protein kinase that serves as a central regulator of cellular metabolism, growth, and survival in response to various signals, such as hormones and nutrients. MTOR operates within two distinct signaling complexes, mTORC1 and mTORC2. mTORC1 is activated to upregulate protein synthesis by phosphorylating regulators of mRNA translation and ribosome synthesis, and phosphorylates and activates proteins like RPS6KB1 and RPS6KB2 to promote protein synthesis. It controls MiT/TFE factors TFEB and TFE3 by mediating their retention and inactivation under nutrient-rich conditions, and it inhibits autophagy by phosphorylating DAP and RUBCNL/Pacer. Additionally, mTORC1 engages in feedback control on growth factor signaling and may influence microtubules through CLIP1 phosphorylation. The mTORC2 complex may regulate cellular processes, including survival and cytoskeletal organization, by phosphorylating AKT1 and regulating the actin cytoskeleton via PRKCA, PXN, and Rho-type guanine nucleotide exchange factors. It also regulates the phosphorylation of SGK1. This supplementary information is collated from multiple sources and compiled automatically.

Alternative names

FRAP, FRAP1, FRAP2, RAFT1, RAPT1, MTOR, Serine/threonine-protein kinase mTOR, FK506-binding protein 12-rapamycin complex-associated protein 1, FKBP12-rapamycin complex-associated protein, Mammalian target of rapamycin, Mechanistic target of rapamycin, Rapamycin and FKBP12 target 1, Rapamycin target protein 1, mTOR

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: Y391
Purification technique: Affinity purification Protein A
Specificity: Expression levels of the target protein vary with sample type and some optimisation may be required.
Epitope: ab32028 reacts with an epitope located in the C terminal region of mTOR.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-mTOR antibody [Y391] (ab32028) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunohistochemistry (IHC-Fr) in Human, Mouse, Rat samples.

What is the molecular weight of mTOR?

Anti-mTOR [Y391] (ab32028) specifically detects a band for mTOR (UniProt: P42345) at a molecular weight of 289kDa.

Trusted by the scientific community

Anti-mTOR [Y391] (ab32028) was first used in a scientific publication in 2006 and has been cited over 230 times in peer-reviewed journals.

Reviewed by scientists

Anti-mTOR [Y391] (ab32028) has over 5 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [Y391] also available for your convenience:
ab32028, Carrier free - ab218525

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

The mammalian target of rapamycin commonly known as mTOR is a serine/threonine kinase known for its role in cellular growth and metabolism. It has a molecular weight of approximately 289 kDa. mTOR is expressed in various tissues throughout the body including muscle adipose tissue and the brain. The protein functions as a central regulator of cell proliferation protein synthesis and nutrient signaling. Often researchers utilize mTOR ELISA or mTOR western blot (mTOR WB) methods and mTOR antibodies to study its expression and activity in various biological contexts.

Biological function summary

MTOR integrates signals from nutrients growth factors and cellular energy status to maintain cellular homeostasis. It forms part of two distinct complexes mTORC1 and mTORC2 which differ in their component proteins and downstream effects. mTORC1 primarily responds to amino acids and regulates protein synthesis through phosphorylation of key substrates like S6K1. On the other hand mTORC2 is important for maintaining cytoskeletal integrity and cell survival highlighting the protein's importance in diverse cellular processes.

Pathways

MTOR plays a pivotal role in the PI3K/AKT/mTOR pathway which governs cell growth proliferation and survival. It also has implications in the regulation of the AMPK pathway which senses cellular energy levels. Through these pathways mTOR interacts with proteins such as AKT and TSC2. The phospho-mTOR specifically the S2448 phospho-mTOR serves as an important functional marker in these signaling cascades linking extracellular signals to downstream cellular responses.

Associated diseases and disorders

MTOR has connections to cancer and neurodegenerative diseases. Its dysregulation often leads to uncontrolled cellular proliferation a hallmark of many cancers. Conditions such as tuberous sclerosis can occur due to mutations in proteins like TSC1 and TSC2 that regulate mTOR activity. In Alzheimer's disease mTOR's role in autophagy and protein synthesis becomes significant as imbalance may contribute to disease progression. Understanding these connections highlights the potential of targeting mTOR pathways therapeutically.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

Western blot - Anti-mTOR antibody [Y391] (ab32028)
Blocking and diluting buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-mTOR antibody [Y391] (ab32028) at 1/2000 dilution
Lane 1: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates in RIPA buffer at 20 µg
Lane 2: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates in HOT buffer at 20 µg
Lane 3: Rat brain lysates in RIPA buffer at 20 µg
Lane 4: Rat brain lysates in HOT buffer at 20 µg
Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg
Lane 6: Human fetal lung lysates at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 290 kDa
Exposure time: 15s
Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10μg)
Lane 2 (+): ab32028 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32028 in HeLa whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
Predicted band size: 289 kDa
Observed band size: 289 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Frozen sections) - Anti-mTOR antibody [Y391] (ab32028)
IHC image of mTOR 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 ab32028, 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.
Western blot - Anti-mTOR antibody [Y391] (ab32028)
MTOR Western blot staining using rabbit Anti-mTOR antibody
Exposure time:
Lane 1: 10 seconds
Lane 2-8: 180 seconds
This antibody detects non-specific bands and high background. It doesn't detect the target band in some mouse and rat tissues.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution
Lane 1: Rat brain tissue lysate at 20 µg
Lane 2: Rat heart tissue lysate at 20 µg
Lane 3: Rat liver tissue lysate at 20 µg
Lane 4: Rat spleen tissue at 20 µg
Lane 5: Mouse brain tissue lysate at 20 µg
Lane 6: Mouse heart tissue lysate at 20 µg
Lane 7: Mouse kidney tissue lysate at 20 µg
Lane 8: Western blot - Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution
Lane 8: Mouse liver tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 289 kDa
Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
ab32028 (purified) at a dilution of 1/100 immunoprecipitating mTOR in rat brain tissue lysate.
Lane 1 (input): Rat brain tissue lysate (10μg)
Lane 2 (+): ab32028 + rat brain tissue lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32028 in rat brain tissue lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028)
Predicted band size: 289 kDa
Observed band size: 289 kDa
Western blot - Anti-mTOR antibody [Y391] (ab32028)
MTOR Western blot staining using rabbit Anti-mTOR antibody
Exposure time:
Lane 1-3: 100 seconds
Lane 4-7: 180 seconds
This antibody detects non-specific bands and high background. It doesn't detect the target band in some mouse and rat tissues.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
All lanes: Western blot - Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HaCaT (Human skin keratinocyte) whole cell lysate at 20 µg
Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: Mouse thymus (8-10 weeks) tissue lysate at 20 µg
Lane 5: Mouse lung (8-10 weeks) tissue lysate at 20 µg
Lane 6: Rat thymus tissue lysate at 20 µg
Lane 7: Rat lung tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 289 kDa
Observed band size: 289 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [Y391] (ab32028)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat testis tissue labelling mTOR with purified ab32028 at a dilution of 1/400. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-mTOR antibody [Y391] (ab32028)
False colour image of Western blot: Anti-mTOR antibody [Y391] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32028 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.
All lanes: Western blot - Anti-mTOR antibody [Y391] (ab32028) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: MTOR [homo] CRISPR-Cas9 edited A549 cell lysate at 20 µg
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: HEK-293 cell lysate at 20 µg
Secondary
Lanes 1 - 4: Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4: Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Observed band size: 250 kDa