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Rabbit Recombinant Monoclonal MTOR antibody. Suitable for IHC-P, IP, WB, IHC-Fr and reacts with Mouse, Rat, Human samples. Cited in 239 publications.


Images

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-PIPFlow CytWBICC/IFIHC-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

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

Rat

Dilution info

1/50 - 1/100

Notes

-

Species

Human

Dilution info

1/50 - 1/100

Notes

-

Expected
Expected

Species

Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Not recommended
Not recommended

Species

Mouse

Dilution info

-

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species

Rat

Dilution info

-

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species

Human

Dilution info

-

Notes

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

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

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

1/200

Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Target data

Function

Serine/threonine protein kinase which is a central regulator of cellular metabolism, growth and survival in response to hormones, growth factors, nutrients, energy and stress signals (PubMed:12087098, PubMed:12150925, PubMed:12150926, PubMed:12231510, PubMed:12718876, PubMed:14651849, PubMed:15268862, PubMed:15467718, PubMed:15545625, PubMed:15718470, PubMed:18497260, PubMed:18762023, PubMed:18925875, PubMed:20516213, PubMed:20537536, PubMed:21659604, PubMed:23429703, PubMed:23429704, PubMed:25799227, PubMed:26018084). MTOR directly or indirectly regulates the phosphorylation of at least 800 proteins. Functions as part of 2 structurally and functionally distinct signaling complexes mTORC1 and mTORC2 (mTOR complex 1 and 2) (PubMed:15268862, PubMed:15467718, PubMed:18925875, PubMed:18497260, PubMed:20516213, PubMed:21576368, PubMed:21659604, PubMed:23429704). Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis (PubMed:12087098, PubMed:12150925, PubMed:12150926, PubMed:12231510, PubMed:12718876, PubMed:14651849, PubMed:15268862, PubMed:15467718, PubMed:15545625, PubMed:15718470, PubMed:18497260, PubMed:18762023, PubMed:18925875, PubMed:20516213, PubMed:20537536, PubMed:21659604, PubMed:23429703, PubMed:23429704, PubMed:25799227, PubMed:26018084). This includes phosphorylation of EIF4EBP1 and release of its inhibition toward the elongation initiation factor 4E (eiF4E) (By similarity). Moreover, phosphorylates and activates RPS6KB1 and RPS6KB2 that promote protein synthesis by modulating the activity of their downstream targets including ribosomal protein S6, eukaryotic translation initiation factor EIF4B, and the inhibitor of translation initiation PDCD4 (PubMed:12150925, PubMed:12087098, PubMed:18925875). This also includes mTORC1 signaling cascade controlling the MiT/TFE factors TFEB and TFE3: in the presence of nutrients, mediates phosphorylation of TFEB and TFE3, promoting their cytosolic retention and inactivation (PubMed:22576015, PubMed:22343943, PubMed:22692423). Upon starvation or lysosomal stress, inhibition of mTORC1 induces dephosphorylation and nuclear translocation of TFEB and TFE3, promoting their transcription factor activity (PubMed:22576015, PubMed:22343943, PubMed:22692423). Stimulates the pyrimidine biosynthesis pathway, both by acute regulation through RPS6KB1-mediated phosphorylation of the biosynthetic enzyme CAD, and delayed regulation, through transcriptional enhancement of the pentose phosphate pathway which produces 5-phosphoribosyl-1-pyrophosphate (PRPP), an allosteric activator of CAD at a later step in synthesis, this function is dependent on the mTORC1 complex (PubMed:23429704, PubMed:23429703). Regulates ribosome synthesis by activating RNA polymerase III-dependent transcription through phosphorylation and inhibition of MAF1 an RNA polymerase III-repressor (PubMed:20516213). In parallel to protein synthesis, also regulates lipid synthesis through SREBF1/SREBP1 and LPIN1 (By similarity). To maintain energy homeostasis mTORC1 may also regulate mitochondrial biogenesis through regulation of PPARGC1A (By similarity). mTORC1 also negatively regulates autophagy through phosphorylation of ULK1 (By similarity). Under nutrient sufficiency, phosphorylates ULK1 at 'Ser-758', disrupting the interaction with AMPK and preventing activation of ULK1 (By similarity). Also prevents autophagy through phosphorylation of the autophagy inhibitor DAP (PubMed:20537536). Also prevents autophagy by phosphorylating RUBCNL/Pacer under nutrient-rich conditions (PubMed:30704899). mTORC1 exerts a feedback control on upstream growth factor signaling that includes phosphorylation and activation of GRB10 a INSR-dependent signaling suppressor (PubMed:21659604). Among other potential targets mTORC1 may phosphorylate CLIP1 and regulate microtubules (PubMed:12231510). As part of the mTORC2 complex MTOR may regulate other cellular processes including survival and organization of the cytoskeleton (PubMed:15268862, PubMed:15467718). Plays a critical role in the phosphorylation at 'Ser-473' of AKT1, a pro-survival effector of phosphoinositide 3-kinase, facilitating its activation by PDK1 (PubMed:15718470). mTORC2 may regulate the actin cytoskeleton, through phosphorylation of PRKCA, PXN and activation of the Rho-type guanine nucleotide exchange factors RHOA and RAC1A or RAC1B (PubMed:15268862). mTORC2 also regulates the phosphorylation of SGK1 at 'Ser-422' (PubMed:18925875). Regulates osteoclastogenesis by adjusting the expression of CEBPB isoforms (By similarity). Plays an important regulatory role in the circadian clock function; regulates period length and rhythm amplitude of the suprachiasmatic nucleus (SCN) and liver clocks (By similarity). Phosphorylates SQSTM1, promoting interaction between SQSTM1 and KEAP1 and subsequent inactivation of the BCR(KEAP1) complex (By similarity).

Alternative names

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Rabbit Recombinant Monoclonal MTOR antibody. Suitable for IHC-P, IP, WB, IHC-Fr and reacts with Mouse, Rat, Human samples. Cited in 239 publications.

Alternative names

Key facts

Isotype

IgG

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
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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

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

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.

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), expandable thumbnail

    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) (AB97051) at 1/20000 dilution

    Predicted band size: 289 kDa

    Observed band size: 290 kDa

    Exposure time: 15s

  • Immunoprecipitation - Anti-mTOR antibody [Y391] (ab32028), expandable thumbnail

    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 (ab172730) instead of ab32028 in HeLa whole cell lysate.

    For western blotting, 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), expandable thumbnail

    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. 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), expandable thumbnail

    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 BONDTM 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), expandable thumbnail

    Western blot - Anti-mTOR antibody [Y391] (ab32028)

    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.

    Lanes 1 - 7: Western blot - Anti-mTOR antibody [Y391] (AB32028)

    Lane 8: 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: Mouse liver tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - 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), expandable thumbnail

    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 (ab172730) instead of ab32028 in rat brain tissue lysate.

    For western blotting, 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), expandable thumbnail

    Western blot - Anti-mTOR antibody [Y391] (ab32028)

    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.

    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) (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), expandable thumbnail

    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. 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), expandable thumbnail

    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. 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), expandable thumbnail

    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] (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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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