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Proteins and peptidesAnti-Ly6g antibody [1A8] - mouse IgG2c (Chimeric)
Low endotoxin, Azide free.
Our first-to-market chimera with mouse IgG2c backbone, this functional antibody specifically depletes neutrophils in vivo for up to 72h.
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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.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
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 |
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. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/50 - 1/100 | Notes - |
Species Human | Dilution info 1/50 - 1/100 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
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. |
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. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat, Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/200 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
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).
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, FRAP1, RAFT1, FRAP, MTOR, FRAP2, RAPT1
Rabbit Recombinant Monoclonal MTOR antibody. Suitable for IHC-P, IP, WB, IHC-Fr and reacts with Mouse, Rat, Human samples. Cited in 239 publications.
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, FRAP1, RAFT1, FRAP, MTOR, FRAP2, RAPT1
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
The exact immunogen used to generate this antibody is proprietary information.
Y391
Affinity purification Protein A
Expression levels of the target protein vary with sample type and some optimisation may be required.
ab32028 reacts with an epitope located in the C terminal region of mTOR.
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Avoid freeze / thaw cycle
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:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our 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.
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
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
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) 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.
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.
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
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
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
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
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) 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) 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.
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
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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