Invitrogen

mTOR Recombinant Rabbit Monoclonal Antibody (SU30-00)

Advanced Verification

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

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mTOR Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of mTOR in paraffin-embedded human breast carcinoma tissue. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with mTOR Recombinant Monoclonal Antibody (Product # MA5-41124) at a dilution of 1:50 for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

mTOR Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of mTOR in paraffin-embedded mouse kidney tissue. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with mTOR Recombinant Monoclonal Antibody (Product # MA5-41124) at a dilution of 1:50 for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

mTOR Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of mTOR in paraffin-embedded mouse testis tissue. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with mTOR Recombinant Monoclonal Antibody (Product # MA5-41124) at a dilution of 1:50 for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

mTOR Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry (Paraffin) analysis of mTOR in paraffin-embedded human kidney tissue. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with mTOR Recombinant Monoclonal Antibody (Product # MA5-41124) at a dilution of 1:50 for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

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 CRISPR637731_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of mTOR was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa Cas9 transduced with mTOR Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with mTOR Recombinant Rabbit Monoclonal Antibody (SU30-00) (Product # MA5-41124, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076) with the iBright™ FL1500 (Product # A44115). A reduced signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific tomTOR.

mTOR Antibody in Western Blot (WB) — Western Blot analysis of mTOR using mTOR Recombinant Monoclonal Antibody (Product # MA5-41124) at a dilution of 1:500 was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody at 1:5,000 dilution was used for 1 hour at room temperature. Lane 1: mouse testis tissue lysate, Lane 2: SiHa cell lysate.

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 mTOR Recombinant Rabbit Monoclonal Antibody (SU30-00) (Product # MA5-41124).

Product Details

MA5-41124

Applications

Tested Dilution

Publications

Western Blot (WB)

1:1,000

-

Immunohistochemistry (Paraffin) (IHC (P))

1:50-1:200

-

Product Specifications

Species Reactivity

Human, Mouse, Rat

Host/Isotype

Rabbit / IgG

Expression System

HEK293 cells

Class

Recombinant Monoclonal

Type

Antibody

Clone

SU30-00

Immunogen

Synthetic peptide within human mTOR aa 2400-2440

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

TBS, pH 7.4, with 0.05% BSA, 40% glycerol

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2898878

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.

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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

negative regulation of protein phosphorylation positive regulation of protein phosphorylation positive regulation of endothelial cell proliferation heart morphogenesis heart valve morphogenesis regulation of glycogen biosynthetic process energy reserve metabolic process 'de novo' pyrimidine nucleobase biosynthetic process DNA repair protein phosphorylation response to stress cell cycle arrest signal transduction germ cell development brain development cell aging response to nutrient long-term memory visual learning post-embryonic development negative regulation of autophagy positive regulation of lamellipodium assembly positive regulation of gene expression positive regulation of myotube differentiation positive regulation of neuron maturation negative regulation of muscle atrophy cell growth macroautophagy negative regulation of macroautophagy phosphorylation peptidyl-serine phosphorylation peptidyl-threonine phosphorylation spinal cord development protein catabolic process positive regulation of actin filament polymerization T cell costimulation negative regulation of protein ubiquitination ruffle organization regulation of myelination cellular response to nutrient levels TOR signaling regulation of fatty acid beta-oxidation regulation of response to food response to insulin regulation of actin cytoskeleton organization social behavior multicellular organism growth growth wound healing response to cocaine regulation of GTPase activity response to amino acid response to morphine regulation of carbohydrate utilization positive regulation of nitric oxide biosynthetic process regulation of osteoclast differentiation positive regulation of translation negative regulation of cell size regulation of protein kinase activity positive regulation of transcription from RNA polymerase III promoter protein autophosphorylation positive regulation of lipid biosynthetic process phosphatidylinositol-mediated signaling mRNA stabilization positive regulation of smooth muscle cell proliferation positive regulation of oligodendrocyte differentiation positive regulation of peptidyl-tyrosine phosphorylation voluntary musculoskeletal movement positive regulation of stress fiber assembly negative regulation of NFAT protein import into nucleus positive regulation of protein kinase B signaling cardiac muscle cell development cardiac muscle contraction maternal process involved in female pregnancy positive regulation of glial cell proliferation positive regulation of dendritic spine development positive regulation of cell growth involved in cardiac muscle cell development cellular response to hypoxia regulation of brown fat cell differentiation regulation of membrane permeability regulation of cellular response to heat positive regulation of neuron death positive regulation of transcription of nuclear large rRNA transcript from RNA polymerase I promoter positive regulation of eating behavior positive regulation of cholangiocyte proliferation positive regulation of sensory perception of pain negative regulation of cholangiocyte apoptotic process positive regulation of granulosa cell proliferation positive regulation of skeletal muscle hypertrophy negative regulation of iodide transmembrane transport double-strand break repair via homologous recombination regulation of carbohydrate metabolic process cell projection organization

Process(es)

RNA polymerase III type 1 promoter DNA binding RNA polymerase III type 2 promoter DNA binding RNA polymerase III type 3 promoter DNA binding TFIIIC-class transcription factor binding protein kinase activity protein serine/threonine kinase activity protein binding ATP binding kinase activity protein kinase binding protein domain specific binding ribosome binding phosphoprotein binding nucleotide binding drug binding transferase activity transferase activity, transferring phosphorus-containing groups phosphotransferase activity, alcohol group as acceptor protein dimerization activity