AB322444

Anti-mTOR antibody [RM1258]

20ul selling size
BOND RX™ Validated
RabMAb
Recombinant
KO Validated
What is this?

Be the first to review this product! Submit a review

(0 Publication)

Rabbit Recombinant Multiclonal MTOR antibody. Suitable for WB, IHC-P, IP and reacts with Human, Mouse, Rat samples.

View 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

11 Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling mTOR with ab322444 at 1/100 (5.17 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human cerebrum. The section was incubated with ab322444 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type 293T (human embryonic kidney epithelial cell) cell pellet (B) MTOR knockout HEK293T cell pellet tissue labeling mTOR with ab322444 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on (A) wild-type 293T cell pellet, no staining on (B) MTOR knockout HEK293T cell pellet. The section was incubated with ab322444 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Supplier Data

Immunoprecipitation - Anti-mTOR antibody [RM1258] (AB322444)

mTOR was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab322444 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322444 at 1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab322444 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322444 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-mTOR antibody [RM1258] (ab322444) at 1/30 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 32s

Supplier Data

Immunoprecipitation - Anti-mTOR antibody [RM1258] (AB322444)

mTOR was immunoprecipitated from 0.35 mg PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate with ab322444 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322444 at 1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 2 : ab322444 IP in PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322444 in PC-12 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-mTOR antibody [RM1258] (ab322444) at 1/30 dilution

All lanes:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 10s

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling mTOR with ab322444 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat cerebrum. The section was incubated with ab322444 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling mTOR with ab322444 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse cerebrum. The section was incubated with ab322444 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

Supplier Data

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-mTOR antibody [RM1258] (ab322444) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 289 kDa,36 kDa

false

Exposure time: 6s

Supplier Data

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab322269 was shown to bind specifically to mTOR. A band was observed at 290 kDa in wild-type 293T cell lysates with whereas no signal observed at this size in mTOR knockout cell line.

The band below 289 kDa may be caused by degradation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-mTOR antibody [RM1258] (ab322444) at 1/1000 dilution

Lane 1:

Wild-type 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

MTOR knockout HEK293T whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 289 kDa,36 kDa

false

Exposure time: 6s

Supplier Data

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The band below 289 kDa may be caused by degradation.

Exposure time : Lane 1 : 6 seconds Lane 2 : 81 seconds

All lanes:

Western blot - Anti-mTOR antibody [RM1258] (ab322444) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 289 kDa

false

Supplier Data

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The band below 289 kDa may be caused by degradation.

All lanes:

Western blot - Anti-mTOR antibody [RM1258] (ab322444) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 289 kDa

false

Exposure time: 6s

Supplier Data

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The band below 289 kDa may be caused by degradation.

All lanes:

Western blot - Anti-mTOR antibody [RM1258] (ab322444) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

false

Exposure time: 6s

Key facts

Host species

Rabbit

Clonality

Multiclonal

Clone number

RM1258

Isotype

IgG

Carrier free

Reacts with

Human, Mouse, Rat

Applications

WB, IP, IHC-P

applications

Immunogen

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

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/100", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/500", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "IP-species-checked": "guaranteed", "IP-species-dilution-info": "", "IP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Rat": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "IHCP-species-checked": "testedAndGuaranteed", "IHCP-species-dilution-info": "1/500", "IHCP-species-notes": " Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.", "IP-species-checked": "testedAndGuaranteed", "IP-species-dilution-info": "1/30", "IP-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "notRecommended", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" } } }

style

left-column

Product details

What are recombinant multiclonals?
Recombinant multiclonals are a mixture of recombinant antibodies co-expressed from a library of heavy and light chains. They offer several advantages including:

- The sensitivity of polyclonal antibodies by recognising multiple epitopes
- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

View our range of recombinant multiclonal antibodies.

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.

style

left-column

Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Preservative: 0.01% Sodium azide Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

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

style

left-column

Supplementary information

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

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.

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.

style

left-column

style

left-column

Product protocols

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

Visit the General protocols
Visit the Troubleshooting

style

left-column

Target data

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.

See full target information MTOR

style

left-column

style

left-column

style

right-column

Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

For licensing inquiries, please contact partnerships@abcam.com

Anti-mTOR antibody [RM1258]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunoprecipitation - Anti-mTOR antibody [RM1258] (AB322444)

Immunoprecipitation - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR antibody [RM1258] (AB322444)

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Western blot - Anti-mTOR antibody [RM1258] (AB322444)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Complementary Products

Assay Kits

Product promise