Invitrogen

mTOR Monoclonal Antibody (215Q18)

Advanced Verification

This Antibody was verified by Knockout to ensure that the antibody binds to the antigen stated.

6 Published Figures

10 References

View all (84) mTOR antibodies

Datasheet

Protocols

Questions & Answers

Tech Support

Datasheet

Protocols

Questions & Answers

Tech Support

Product Image Gallery

mTOR Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescent analysis of mTOR was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with mTOR Mouse monoclonal Antibody (Product # AHO1232) at 2 µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

mTOR Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of mTOR showing staining in the cytoplasm of paraffin-embedded human testis tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a mTOR Mouse Monoclonal Antibody (clone 215Q18) (Product # AHO1232) diluted in 3% BSA-PBS at a dilution of 1:50 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

mTOR Antibody in Western Blot (WB) — CRISPR-Cas9 mediated genome editing ofmTOR (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR814789_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of mTOR was performed by loading 60 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Cas9 cells transduced with mTOR Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-mTOR Monoclonal Antibody (215Q18) (Product # AHO1232) using 1 µg/mL dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:5000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line G2confirms that antibody is specific tomTOR (Fig (b)).

mTOR Antibody in Immunohistochemistry (IHC) — Figure 1 Knockdown of YAP1 and mTOR inhibited the progression of bladder cancer. (A and B) RT-PCR and western blot assays were performed to test the expression of YAP1 and mTOR in bladder cancer and normal bladder tissues ( *** P<0.001). (C) Immunohistochemistry was used to evaluate the expression patterns of the YAP1 and mTOR proteins in three matched pairs of bladder cancer and normal bladder tissues (magnification, x200). (D and E) Immunohistochemistry scores of mTOR and YAP1 staining in 20 paired bladder cancer and adjacent normal tissues. (F and G) RT-PCR was used to analyze the knockdown efficien-cies of si-YAP1 and si-mTOR ( * P<0.05, ** P<0.01). (H and I) HT-1376 and J82 cells were transfected with si-NC, si-YAP1, si-mTOR or si-YAP1 + si-mTOR for 0, 24, 48, 72 and 96 h; then, CCK-8 assay was performed to evaluate cell proliferation. (J) HT-1376 and J82 cells were transfected with si-NC, si-YAP1, si-mTOR or si-YAP1 + si-mTOR for 48 h; then, cells were collected for flow cytometry assay to detect cell apoptosis. (K) Western blotting was performed to detect the expression of apoptosis-related proteins, such as caspase 3/9 and cleaved-caspase3/9 after 48 h of cell transfection (H-K, vs. si-NC group, * P<0.05; vs. si-YAP1 group, # P<0.05; NC, negative control). YAP1, Yes-associated protein 1; mTOR, mammalian target of rapamycin; RT-PCR, reverse transcription-polymerase chain reaction.

mTOR Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 2 Crosstalk between the YAP1 and mTOR proteins. (A and B) HT-1376 and J82 cells were transfected with OE-mTOR and OE-NC; then cells were harvested and subjected to RT-PCR and western blot assays to determine the expression of mTOR at the mRNA and protein levels, respectively. (C-F) RT-PCR and western blot assays were performed to determine the mRNA and protein expression of YAP1 and TEAD after 48 h of HT-1376 and J82 cell transfection with OE-mTOR or OE-NC. (G) Immunofluorescence assay was performed to evaluate the effects of subcellular location of the YAP1 protein. (H) Statistical analysis of the fluorescence intensity of the YAP1 protein ( * P<0.05, ** P<0.01; NC, negative control). YAP1, Yes-associated protein 1; mTOR, mammalian target of rapamycin; RT-PCR, reverse transcription-polymerase chain reaction; TEAD, TEA domain transcription factor.

mTOR Antibody — Antibody specificity was demonstrated by CRISPR-Cas9 mediated knockout of target protein. A reduced signal was observed for target protein in HeLa Cas9 cell line transduced with mTOR Lentiviral sgRNA compared to control cell line using Anti-mTOR Monoclonal Antibody (215Q18) (Product # AHO1232). {KO}

Product Details

AHO1232

Applications	Tested Dilution	Publications
Western Blot (WB)	Assay-dependent	View 5 publications 5 publications
Immunohistochemistry (IHC)	-	View 2 publications 2 publications
Immunohistochemistry (Paraffin) (IHC (P))	1:10-1:100	-
Immunocytochemistry (ICC/IF)	1:100-1:500	View 2 publications 2 publications
Immunoprecipitation (IP)	-	View 1 publication 1 publication

Product Specifications
Species Reactivity	Human, Mouse, Rat
Published species	Human, Mouse, Rat
Host/Isotype	Mouse / IgG2b, kappa
Class	Monoclonal
Type	Antibody
Clone	215Q18
Immunogen	Recombinant fragment of human mTOR expressed in E. coli.
Conjugate	Unconjugated
Form	Liquid
Concentration	0.5 mg/mL
Purification	Affinity chromatography
Storage buffer	PBS, pH 7.2, with 1% BSA
Contains	0.1% sodium azide
Storage conditions	-20°C
RRID	AB_2536329

Target Information

Mammalian target of rapamycin (mTOR) is a serine/threonine kinase that plays a key role in cell growth, cell proliferation, and protein synthesis. mTOR mediates phosphoinositide 3-kinase and Akt/PKB signaling, resulting in phosphorylation of 4EBP1, and initiation of mRNA translation. A second pathway involves regulation of ribosomal S6 kinase, which affects ribosome biogenesis and translation elongation. Mutations affecting the gene results in Smith-Kingsmore syndrome.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Anti-inflammatory and Antitumor Activity of a Triple Therapy for a Colitis-Related Colorectal Cancer.

Journal of Cancer

Figueroa-González G,García-Castillo V,Coronel-Hernández J,López-Urrutia E,León-Cabrera S,Arias-Romero LE,Terrazas LI,Rodríguez-Sosa M,Campos-Parra AD,Zúñiga-Calzada E,Lopez-Camarillo C,Morales-González F,Jacobo-Herrera NJ,Pérez-Plasencia C

AHO1232 was used in western blot to discuss targeting mTOR-signaling and lactate dehydrogenase A as a strategy to treat colorectal cancer

Fri Apr 08 00:00:00 EDT 2022

Discovery of a novel potentially transforming somatic mutation in CSF2RB gene in breast cancer.

Cancer medicine

Rashid M,Ali R,Almuzzaini B,Song H,AlHallaj A,Abdulkarim AA,Mohamed Baz O,Al Zahrani H,Mustafa Sabeena M,Alharbi W,Hussein M,Boudjelal M

Published figure using mTOR monoclonal antibody (Product # AHO1232) in Western Blot

Mon Nov 01 00:00:00 EDT 2021

Isolation and Establishment of a Highly Proliferative, Cancer Stem Cell-Like, and Naturally Immortalized Triple-Negative Breast Cancer Cell Line, KAIMRC2.

Cells

Ali R,Al Zahrani H,Barhoumi T,Alhallaj A,Mashhour A,Alshammari MA,Alshawakir YA,Baz O,Alanazi AH,Khan AL,Al Nikhli H,Al Balwi MA,Al Riyees L,Boudjelal M

AHO1232 was used in Western Blot to report the establishment of a naturally immortalized highly tumorigenic and triple-negative breast cancer cell line, KAIMRC2.

Mon May 24 00:00:00 EDT 2021

Isolation and Establishment of a Highly Proliferative, Cancer Stem Cell-Like, and Naturally Immortalized Triple-Negative Breast Cancer Cell Line, KAIMRC2.

Cells

Ali R,Al Zahrani H,Barhoumi T,Alhallaj A,Mashhour A,Alshammari MA,Alshawakir YA,Baz O,Alanazi AH,Khan AL,Al Nikhli H,Al Balwi MA,Al Riyees L,Boudjelal M

AHO1232 was used in Western Blot to report the establishment of a naturally immortalized highly tumorigenic and triple-negative breast cancer cell line, KAIMRC2.

Mon May 24 00:00:00 EDT 2021

UPF2 leads to degradation of dendritically targeted mRNAs to regulate synaptic plasticity and cognitive function.

Molecular psychiatry

Notaras M,Allen M,Longo F,Volk N,Toth M,Li Jeon N,Klann E,Colak D

AHO1232 was used in Immunohistochemistry to establish that UPF2 regulates synaptic plasticity, cognition, and local protein synthesis in dendrites, providing fundamental insight into the neuron-specific function of NMD within the brain.

Tue Dec 01 00:00:00 EST 2020

Interaction of YAP1 and mTOR promotes bladder cancer progression.

International journal of oncology

Xu M,Gu M,Zhou J,Da J,Wang Z

Published figure using mTOR monoclonal antibody (Product # AHO1232) in Immunofluorescence

Wed Jan 01 00:00:00 EST 2020

Interaction of YAP1 and mTOR promotes bladder cancer progression.

International journal of oncology

Xu M,Gu M,Zhou J,Da J,Wang Z

Published figure using mTOR monoclonal antibody (Product # AHO1232) in Immunofluorescence

Wed Jan 01 00:00:00 EST 2020

Interaction of YAP1 and mTOR promotes bladder cancer progression.

International journal of oncology

Xu M,Gu M,Zhou J,Da J,Wang Z

Published figure using mTOR monoclonal antibody (Product # AHO1232) in Immunofluorescence

Wed Jan 01 00:00:00 EST 2020

Interaction of YAP1 and mTOR promotes bladder cancer progression.

International journal of oncology

Xu M,Gu M,Zhou J,Da J,Wang Z

Published figure using mTOR monoclonal antibody (Product # AHO1232) in Immunofluorescence

Wed Jan 01 00:00:00 EST 2020

A Stat6/Pten Axis Links Regulatory T Cells with Adipose Tissue Function.

Cell metabolism

Kälin S,Becker M,Ott VB,Serr I,Hosp F,Mollah MMH,Keipert S,Lamp D,Rohner-Jeanrenaud F,Flynn VK,Scherm MG,Nascimento LFR,Gerlach K,Popp V,Dietzen S,Bopp T,Krishnamurthy P,Kaplan MH,Serrano M,Woods SC,Tripal P,Palmisano R,Jastroch M,Blüher M,Wolfrum C,Weigmann B,Ziegler AG,Mann M,Tschöp MH,Daniel C

AHO1232 was used in Immunocytochemistry-immunoflourescence to study the link between adipose tissue function and regulatory T cells.

Tue Sep 05 00:00:00 EDT 2017

Rapamycin reveals an mTOR-independent repression of Kv1.1 expression during epileptogenesis.

Neurobiology of disease

Sosanya NM,Brager DH,Wolfe S,Niere F,Raab-Graham KF

AHO1232 was used in western blot to study mTOR, microRNA miR-129-5p, and Kv1.1 in epilepsy

Thu Jan 01 00:00:00 EST 2015

Bioinformatics

Protein Aliases: angiopoietin-like factor CDT6; FK506 binding protein 12-rapamycin associated protein 1; FK506 binding protein 12-rapamycin associated protein 2; FK506-binding protein 12-rapamycin complex-associated protein 1; FKBP-rapamycin associated protein; FKBP-rapamycin associated protein (FRAP); FKBP-rapamycin-associated protein FRAP; FKBP12-rapamycin complex-associated protein; FKBP12-rapamycin complex-associated protein 1; m-TOR; Mammalian target of rapamycin; Mechanistic target of rapamycin; mechanistic target of rapamycin (serine/threonine kinase); mTOR; mTORC1; Rapamycin and FKBP12 target 1; rapamycin and FKBP12 target-1 protein; rapamycin associated protein FRAP2; Rapamycin target protein 1; RAPT1; Serine/threonine-protein kinase mTOR

Gene Aliases: 2610315D21Rik; AI327068; flat; FRAP; FRAP1; FRAP2; MTOR; RAFT1; RAPT1; SKS

UniProt ID: (Human) P42345, (Mouse) Q9JLN9, (Rat) P42346

Entrez Gene ID: (Human) 2475, (Mouse) 56717, (Rat) 56718

Molecular Function: non-receptor serine/threonine protein kinase protein modifying enzyme