Anti-mTOR 抗体 (ab2833)

Reviews (1)Q&A (5)References (11)
Western blot - Anti-mTOR antibody (ab2833)
  • Immunoprecipitation - Anti-mTOR antibody (ab2833)

Key features and details

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

製品の概要

  • 製品名

    Anti-mTOR antibody
    mTOR 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to mTOR

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: IP, WBmore details

  • 種交差性

    交差種: Human
    交差が予測される動物種: Mouse, Rat

  • 免疫原

    Synthetic peptide within Human mTOR aa 200-300. The exact immunogen sequence used to generate this antibody is proprietary information. If additional detail on the immunogen is needed to determine the suitability of the antibody for your needs, please contact our Scientific Support team to discuss your requirements. NP_004949.1 (GeneID 2475).
    Database link: P42345

  • ポジティブ･コントロール

    • WB: HeLa and HEK-293T whole cell lysate. IP: mTOR IP in HeLa whole cell lysate.

  • 特記事項

    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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab2833の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IP
Use at 2-10 µg/mg of lysate.
WB (1)
1/1000 - 1/5000. Detects a band of approximately 289 kDa (predicted molecular weight: 289 kDa).
特記事項
IP
Use at 2-10 µg/mg of lysate.
WB
1/1000 - 1/5000. Detects a band of approximately 289 kDa (predicted molecular weight: 289 kDa).

ターゲット情報

  • 機能

    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'.

  • 組織特異性

    Expressed in numerous tissues, with highest levels in testis.

  • 配列類似性

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

  • 翻訳後修飾

    Autophosphorylated; when part of mTORC1 or mTORC2.

  • 細胞内局在

    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.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-mTOR antibody (ab2833)
    Western blot - Anti-mTOR antibody (ab2833)
    All lanes : Anti-mTOR antibody (ab2833) at 0.4 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

    Lysates/proteins at 50 µg per lane.

    Predicted band size: 289 kDa


    Exposure time: 30 seconds


    Lysates prepared using NETN lysis buffer.

  • Immunoprecipitation - Anti-mTOR antibody (ab2833)
    Immunoprecipitation - Anti-mTOR antibody (ab2833)

    Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer.
    Lane 1: ab2833 used for IP at 6 µg per reaction.
    Lane 2: Control IgG.
    For blotting immunoprecipitated mTOR, ab2833 was used at 1 µg/ml.
    Detection: Chemiluminescence with an exposure time of 30 seconds.

参考文献 (11)

ab2833 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab2833 は 11 報の論文で使用されています。

  • Yuan X  et al. NUCKS1 is a novel regulator of milk synthesis in and proliferation of mammary epithelial cells via the mTOR signaling pathway. J Cell Physiol N/A:N/A (2019). PubMed: 30710349
  • Yu Y  et al. U2AF65 enhances milk synthesis and growth of bovine mammary epithelial cells by positively regulating the mTOR-SREBP-1c signalling pathway. Cell Biochem Funct 37:93-101 (2019). PubMed: 30773658
  • Liu Y  et al. GRP78 regulates milk biosynthesis and the proliferation of bovinemammaryepithelial cells through the mTOR signaling pathway. Cell Mol Biol Lett 24:57 (2019). PubMed: 31660059
  • Li B  et al. Septin6 regulates cell growth and casein synthesis in dairy cow mammary epithelial cells via mTORC1 pathway. J Dairy Res 86:181-187 (2019). PubMed: 31122298
  • Li P  et al. FABP5 is a critical regulator of methionine- and estrogen-induced SREBP-1c gene expression in bovine mammary epithelial cells. J Cell Physiol 234:537-549 (2018). PubMed: 30071125
View all Publications for this product

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1-6 of 6 Abreviews or Q&A

Western blot abreview for Anti-mTOR antibody

 Average
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Keratinocyte)
Loading amount
15 µg
Specification
Keratinocyte
Gel Running Conditions
Reduced Denaturing (10% gel)
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Read More

Prof. Paul Townsend

Verified customer

投稿 Jun 26 2008

Question

I am looking for an mTOR antibody produced from an immunogen peptide with close homology to the honey bee TOR homolog. For some of your relevant antibodies the exact sequence is not provided. Could you please send me the immunogen peptide sequence for: ab2833: Synthetic peptide (Human) which represents a portion of human Mammalian Target of Rapamycin encoded in part by exons 5 and 6. ab25880: Synthetic 15 amino acid peptide from the amino terminus of human TOR Thanks so much for your help. I hope one of your antibodies will be a good match.

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Answer

Thank you for your enquiry. Unfortunately the immunogen sequences for both ab2833 and ab25880 are proprietary. I can however tell you that with regards ab25880; based on an alignment with NCBI accession number XP_625130 [**- *:* * ***: ] the honey bee shows a degree of similarity making it possible that this antibody will cross-react with its homolog. The immunogen sequence for ab2833 is disimilar to fruit fly or mosquito and therefore is unlikely to cross react with honey bee. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Thank you for your email! We loaded 20 µg as instructed by the specification sheet of the lysate but you are right in pointing out that indeed 50 µg was loaded when using the ab 2833 antibody (we are using both ab 2833 and 2732). Although I am uncertain whether we have 50 µg left of the lysate (50µg is half of what was supplied!) we will load the remaining lysate on the next gel; I have serious doubts that it will make difference however since there was no band at the correct mol weight region.... We are working with the placenta and we have obtained single strong, distinct bands at 230-240 kDa in both human and rat placenta. If we were able to verify that this band indeed is mTOR I believe that our gels may be something for your specification sheet in the future. A positive control (liver lysate) would have been a little help however access to the antigenic peptide would be better. Are there antigenic peptides available for ab 2833 and 2732? Our preliminary data on mTOR in the placenta and in various pregnancy complications has made us quite excited and I will order some related products (1093 and 5454) tonight

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Answer

Thank you for your email. How did the lysate work out for you? I'm glad to hear that you are getting results with your experimental samples but know how important it is to verify what the band is!

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Question

Can you tell me if it should recognize the following section of the FRAP protein (The rapamycin binding domain): SRILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMK SGNVKDLLQAWDLYYHVFRRISK

Read More
Answer

I'm soory but all the information that we have regarding the immunogen is that it is a synthetic peptide (Human) which represents a portion of human Mammalian Target of Rapamycinencoded in part by exons 5 and 6. If you have any more questions please contact us again.

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Question

What is the peptide sequence of the antigen used to raise the mTOR antibody : ab2833?

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Answer

The sequence of the peptide used is proprietary.

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Question

One of the antibodies we have recently purchased from you is anti- mTOR ab2833 which, according to the data sheet, has been tested successfully in western blot. We have a long experience using Western; however so far we have not been able to produce a 235 kDa band shown in your data sheet, instead we obtain a prominent band at half that size. We have so far only used denaturating conditions so my question is whether the 235 kDa band in your data sheet was obtained under denaturating or non-denaturating conditions?? Any other ideas to optimize the western with this particular antibody?

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Answer

Yes, denaturing conditions were used when this antibody was tested in WB. Consider using a lower percentage gel and long transfer times at low voltage to assure transfer of mTor.

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