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Anti-mTOR抗体(ab2732)

  • Datasheet
  • SDS
Reviews (14)Q&A (16)References (349)

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Western blot - Anti-mTOR antibody (ab2732)
  • Western blot - Anti-mTOR antibody (ab2732)
  • Immunoprecipitation - Anti-mTOR antibody (ab2732)
  • Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)
  • Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)

Key features and details

  • Rabbit polyclonal to mTOR
  • Suitable for: ICC/IF, IP, WB
  • Reacts with: Rat, Human
  • Isotype: IgG

选择批间可重复性更高的重组抗体

Product image
Anti-mTOR antibody [Y391] (ab32028)
  • 研究可靠 —— 各批次间结果一致且可重复
  • 长期批量供应 —— 采用重组技术,可实现快速生产
  • 首次实验即可成功 —— 经过大量验证确认了特异性
  • 符合伦理标准 —— 产品不含动物成分

概述

  • 产品名称

    Anti-mTOR抗体
    参阅全部 mTOR 一抗
  • 描述

    兔多克隆抗体to mTOR
  • 宿主

    Rabbit
  • 经测试应用

    适用于: ICC/IF, IP, WBmore details
  • 种属反应性

    与反应: Rat, Human
    预测可用于: Mouse
  • 免疫原

    Synthetic peptide within Human mTOR aa 200-250. The exact sequence is proprietary.
    Database link: P42345
    (Peptide available as ab39393)

  • 阳性对照

    • ICC/IF: HepG2 cells. L6 myotubes. IP: HeLa lysates. WB: HeLa, HEK-293T, Jurkat-HepG2 and LNCaP whole cell lysate.
  • 常规说明

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 存储溶液

    pH: 7
    Preservative: 0.1% Sodium azide
    Constituents: 0.021% PBS, 1.764% Sodium citrate, 1.815% Tris
  • Concentration information loading...
  • 纯化说明

    Affinity purified using the immunising peptideimmobilized on solid support.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • DNA Damage Recognition
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相关产品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant Human mTOR protein (ab114179)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab2732于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
ICC/IF (1)
1/100.
IP
Use at 2-10 µg/mg of lysate.
WB (7)
1/2000 - 1/10000. Predicted molecular weight: 289 kDa.
说明
ICC/IF
1/100.
IP
Use at 2-10 µg/mg of lysate.
WB
1/2000 - 1/10000. Predicted molecular weight: 289 kDa.

靶标

  • 功能

    Kinase subunit of both mTORC1 and mTORC2, which regulates cell growth and survival in response to nutrient and hormonal signals. mTORC1 is activated in response to growth factors or amino-acids. Growth factor-stimulated mTORC1 activation involves AKT1-mediated phosphorylation of TSC1-TSC2, which leads to the activation of the RHEB GTPase that potently activates the protein kinase activity of mTORC1. Amino-acid-signaling to mTORC1 requires its relocalization to the lysosomes mediated by the Ragulator complex and the Rag GTPases. Activated mTORC1 up-regulates protein synthesis by phosphorylating key regulators of mRNA translation and ribosome synthesis. mTORC1 phosphorylates EIF4EBP1 and releases it from inhibiting the elongation initiation factor 4E (eiF4E). mTORC1 phosphorylates and activates S6K1 at 'Thr-421', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Phosphorylates MAF1 leading to attenuation of its RNA polymerase III-repressive function. mTORC2 is also activated by growth. factors, but seems to be nutrient-insensitive. mTORC2 seems to function upstream of Rho GTPases to regulate the actin cytoskeleton, probably by activating one or more Rho-type guanine nucleotide exchange factors. mTORC2 promotes the serum-induced formation of stress-fibers or F-actin. mTORC2 plays a critical role in AKT1 'Ser-473' phosphorylation, which may facilitate the phosphorylation of the activation loop of AKT1 on 'Thr-308' by PDK1 which is a prerequisite for full activation. mTORC2 regulates the phosphorylation of SGK1 at 'Ser-422'. mTORC2 also modulates the phosphorylation of PRKCA on 'Ser-657'.
  • 组织特异性

    Expressed in numerous tissues, with highest levels in testis.
  • 序列相似性

    Belongs to the PI3/PI4-kinase family.
    Contains 1 FAT domain.
    Contains 1 FATC domain.
    Contains 7 HEAT repeats.
    Contains 1 PI3K/PI4K domain.
  • 翻译后修饰

    Autophosphorylated; when part of mTORC1 or mTORC2.
  • 细胞定位

    Endoplasmic reticulum membrane. Golgi apparatus membrane. Mitochondrion outer membrane. Lysosome. Cytoplasm. Nucleus > PML body. Shuttles between cytoplasm and nucleus. Accumulates in the nucleus in response to hypoxia (By similarity). Targeting to lysosomes depends on amino acid availability and RRAGA and RRAGB.
  • Target information above from: UniProt accession P42345 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 2475 Human
    • Entrez Gene: 56717 Mouse
    • Entrez Gene: 56718 Rat
    • Omim: 601231 Human
    • SwissProt: P42345 Human
    • SwissProt: Q9JLN9 Mouse
    • SwissProt: P42346 Rat
    • Unigene: 338207 Human
    • Unigene: 21158 Mouse
    • Unigene: 11008 Rat
    see all
  • 别名

    • dJ576K7.1 (FK506 binding protein 12 rapamycin associated protein 1) antibody
    • FK506 binding protein 12 rapamycin associated protein 1 antibody
    • FK506 binding protein 12 rapamycin associated protein 2 antibody
    • FK506 binding protein 12 rapamycin complex associated protein 1 antibody
    • FK506-binding protein 12-rapamycin complex-associated protein 1 antibody
    • FKBP rapamycin associated protein antibody
    • FKBP12 rapamycin complex associated protein antibody
    • FKBP12-rapamycin complex-associated protein 1 antibody
    • FKBP12-rapamycin complex-associated protein antibody
    • FLJ44809 antibody
    • FRAP antibody
    • FRAP1 antibody
    • FRAP2 antibody
    • Mammalian target of rapamycin antibody
    • Mechanistic target of rapamycin antibody
    • mTOR antibody
    • MTOR_HUMAN antibody
    • OTTHUMP00000001983 antibody
    • RAFT1 antibody
    • Rapamycin and FKBP12 target 1 antibody
    • Rapamycin associated protein FRAP2 antibody
    • Rapamycin target protein 1 antibody
    • Rapamycin target protein antibody
    • RAPT1 antibody
    • Serine/threonine-protein kinase mTOR antibody
    see all

图片

  • Western blot - Anti-mTOR antibody (ab2732)
    Western blot - Anti-mTOR antibody (ab2732)
    All lanes : Anti-mTOR antibody (ab2732) at 1/2000 dilution

    Lane 1 : Wild-type A549 cell lysate
    Lane 2 : MTOR [homo] CRISPR-Cas9 edited A549 cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : HEK-293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 289 kDa
    Observed band size: 250 kDa why is the actual band size different from the predicted?



    False colour image of Western blot: Anti-mTOR antibody staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab2732 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.

  • Western blot - Anti-mTOR antibody (ab2732)
    Western blot - Anti-mTOR antibody (ab2732)
    All lanes : Anti-mTOR antibody (ab2732) at 0.1 µg/ml

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
    Lane 5 : LNCaP (Human prostate cancer cell line) whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Predicted band size: 289 kDa


    Exposure time: 3 minutes


    Lysates prepared using NETN lysis buffer. 

  • Immunoprecipitation - Anti-mTOR antibody (ab2732)
    Immunoprecipitation - Anti-mTOR antibody (ab2732)

    mTOR was immunoprecipitated from HeLa cell lysate (1.0 mg per IP reaction; 20% of IP loaded) with ab2732 at 3 µg per reaction. mTOR was also immunoprecipitated by ab2833. Western blot was performed from the immunoprecipitate with ab2732 at 1 µg/ml.

    Lane 1: ab2833 IP in HeLa whole cell lysate.
    Lane 2: ab2732 IP in HeLa whole cell lysate. 
    Lane 3: Control IgG IP in HeLa  whole cell lysate.

    Detection: Chemiluminescence

  • Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)
    Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)
    ICC/IF image of ab2732 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2732, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)
    Immunocytochemistry/ Immunofluorescence - Anti-mTOR antibody (ab2732)
    ab2732 at a 1:100 dilution confocally staining mTOR (red) in L6 myotubes, alongside a nuclear antigen antibody (green).    

实验方案

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (349)

发表研究结果有使用 ab2732?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab2732 被引用在 349 文献中.

  • Lu Y  et al. Impairment of Autophagy Mediates the Uric-Acid-Induced Phenotypic Transformation of Vascular Smooth Muscle Cells. Pharmacology 109:34-42 (2024). PubMed: 38011839
  • Ning S  et al. MicroRNA-494 regulates high glucose-induced cardiomyocyte apoptosis and autophagy by PI3K/AKT/mTOR signalling pathway. ESC Heart Fail 10:1401-1411 (2023). PubMed: 36772911
  • Wang Y  et al. Rumen-protected glucose stimulates the secretion of reproductive hormones and the mTOR/AKT signaling pathway in the ovaries of early postpartum. Sci Rep 13:2940 (2023). PubMed: 36808140
  • Zhu S  et al. DNA methylation and miR-92a-3p-mediated repression of HIP1R promotes pancreatic cancer progression by activating the PI3K/AKT pathway. J Cell Mol Med 27:788-802 (2023). PubMed: 36811277
  • Ibrahim YH  et al. An affinity tool for the isolation of endogenous active mTORC1 from various cellular sources. J Biol Chem 299:104644 (2023). PubMed: 36965617
View all Publications for this product

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Western blot abreview for Anti-mTOR antibody

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Application
Western blot
Sample
Human Cell lysate - whole cell (PC3 Prostate cancer cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
PC3 Prostate cancer cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jun 12 2019

Western blot abreview for Anti-mTOR antibody

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abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HT1080 cells)
Loading amount
10 µg
Specification
HT1080 cells
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Nov 18 2009

Western blot abreview for Anti-mTOR antibody

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abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (L6 cell line)
Loading amount
10 µg
Specification
L6 cell line
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Nov 18 2009

Western blot abreview for Anti-mTOR antibody

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Application
Western blot
Sample
Human Tissue lysate - whole (Human Penile Tissue)
Loading amount
60 µg
Specification
Human Penile Tissue
Gel Running Conditions
Reduced Denaturing (4-20% criterion gel)
Blocking step
Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jul 30 2008

Western blot abreview for Anti-mTOR antibody

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Western blot
Sample
Mouse Tissue lysate - whole (Cerebellum)
Loading amount
30 µg
Specification
Cerebellum
Gel Running Conditions
Reduced Denaturing (4-20 % gel)
Blocking step
Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Jul 15 2008

Western blot abreview for Anti-mTOR antibody

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Western blot
Sample
Rat Tissue lysate - whole (Penile Tissue)
Loading amount
60 µg
Specification
Penile Tissue
Gel Running Conditions
Reduced Denaturing (7.5 % criterion gel)
Blocking step
Starting Block T20 (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 May 09 2008

Western blot abreview for Anti-mTOR antibody

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Western blot
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Human Tissue lysate - whole (Placenta)
Loading amount
20 µg
Specification
Placenta
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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提交于 Sep 21 2007

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