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Anti-mTOR (phospho S2448) 抗体 (ab1093)

Submit a review Q&A (1)References (23)
Western blot - Anti-mTOR (phospho S2448) antibody (ab1093)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody (ab1093)

Key features and details

  • Rabbit polyclonal to mTOR (phospho S2448)
  • Suitable for: IHC-P, WB, ELISA
  • Reacts with: Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-mTOR (phospho S2448) antibody [EPR426(2)] (ab109268)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-mTOR (phospho S2448) antibody
    mTOR 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to mTOR (phospho S2448)

  • 由来種

    Rabbit

  • 特異性

    Reactivity occurs with phosphorylated mTOR from human derived tissues and cells. Reactivity against mTOR from other species has not been determined, however, reactivity with mouse and rat is suggested based on protein sequence homologies.

  • アプリケーション

    適用あり: IHC-P, WB, ELISAmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide corresponding to Human mTOR aa 2400-2500 (C terminal) (phospho S2448).

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • WB: IGF-1 treated serum starved 293T whole cell lysate. IHC-P: Human kidney tissue, proximal convoluted tubules.

  • 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ab1093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
IHC-P
Use a concentration of 5 µg/ml.
WB
1/1000. Detects a band of approximately 250 kDa (predicted molecular weight: 288 kDa).
ELISA
1/10000 - 1/100000.
特記事項
IHC-P
Use a concentration of 5 µg/ml.
WB
1/1000. Detects a band of approximately 250 kDa (predicted molecular weight: 288 kDa).
ELISA
1/10000 - 1/100000.

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

    • 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 (phospho S2448) antibody (ab1093)
      Western blot - Anti-mTOR (phospho S2448) antibody (ab1093)
      All lanes : Anti-mTOR (phospho S2448) antibody (ab1093) at 1/1000 dilution

      Lane 1 : 24hr serum starved 293T whole cell lysate
      Lane 2 : 24hr serum starved 293T whole cell lysate, treated with IGF-1 (100 ng/ml) for 20 mins prior to harvest

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : HRP Goat-anti-Rabbit IgG at 1/2000 dilution

      Predicted band size: 288 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody (ab1093)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mTOR (phospho S2448) antibody (ab1093)

      Formalin-fixed, paraffin-embedded human kidney tissue, (proximal convulted tubules), labeling mTOR using ab1093 at 5 µg/ml (red). Counterstain is hematoxylin.

    参考文献 (23)

    ab1093 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。

    ab1093 は 23 報の論文で使用されています。

    • Liu M  et al. USP18 contributes to the proliferation and migration of ovarian cancer cells by regulating the AKT/mTOR signaling pathway. Acta Biochim Pol 69:417-422 (2022). PubMed: 35696596
    • Liu L  et al. Ubiquitin-specific protease 8 inhibits lipopolysaccharide-triggered pyroptosis of human bronchial epithelial cells by regulating PI3K/AKT and NF-?B pathways. Allergol Immunopathol (Madr) 50:96-103 (2022). PubMed: 35257552
    • Chen Y  et al. Tripartite motif containing 28 (TRIM28) promotes the growth and migration of endometrial carcinoma cells by regulating the AKT/mTOR signaling pathway. Gen Physiol Biophys 40:245-252 (2021). PubMed: 34100380
    • Ortega MA  et al. Abnormal proinflammatory and stressor environmental with increased the regulatory cellular IGF-1/PAPP-A/STC and Wnt-1/β-Catenin canonical pathway in placenta of women with Chronic venous Disease during Pregnancy. Int J Med Sci 18:2814-2827 (2021). PubMed: 34220309
    • Seidel M  et al. Identification of an amino-terminus determinant critical for ryanodine receptor/Ca2+ release channel function. Cardiovasc Res 117:780-791 (2021). PubMed: 32077934
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    Question

    BATCH NUMBER 125112 ORDER NUMBER 94471 DESCRIPTION OF THE PROBLEM No signal at all SAMPLE WCL isolated from cultured HepG2 , HCT116, DLD1, SUV480 and HT29 cells PRIMARY ANTIBODY Dilution: 1:1000 Dilution buffer: TBS-Tween 20(0.1%) with 5% BSA incubation: overnight at 4 degrees Waching step: wash with TBS-Tween 20(0.1%) , 5min x 3 DETECTION METHOD ECL plus system POSITIVE AND NEGATIVE CONTROLS USED No, the customer didn't use both positive and negative controls. ANTIBODY STORAGE CONDITIONS It was stored at -20 degrees. SAMPLE PREPARATION Lysis buffer: CelLytic-M from SIGMA (cat# C2978) Protease inhibitors: protease inhibitor coacktai tablets from Roche (cat# 11697 498001) Loading buffer: TrisGlycine-SDS sample buffer (2x) from Invitrogen Heating sample: for 5 minutes at 95 degrees AMOUNT OF PROTEIN LOADED approximately 20ug each ELECTROPHORESIS/GEL CONDITIONS 4-12% Tris-Glycine Gel (1.0mm thick) from Invitrogen TRANSFER AND BLOCKING CONDITIONS Blocking buffer: TBS-Tween 20 (0.1%) Blocking agent: 5% non-fat dry milk incubation : for 1 hour at room temp SECONDARY ANTIBODY 2nd Ab: Anti rabbit IgG-HRP conjugated Dilution: 1:10000 Incubation: for 1 hour at room temp Wash step: TBS-Tween 20(0.1%), 5min x 3 HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 ADDITIONAL NOTES Would you please let me know how we should advise her about this matter?

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    Answer

    I'm sorry to hear one of your customers is experiencing a problem with ab1093. We have not tested this antibody in cultured HepG2 , HCT116, DLD1, SUV480 and HT29 cells and I think the customer's problem is due to the fact that these cells contain low levels of the phosphorylated protein, undetectable by WB with ab1093. For example, a paper by Paajarvi et al (FASEB J. 2005 Mar;19(3):476-8. Epub 2004 Dec 29.) shows that HepG2 cells (even serum starved) have very low levels of basal phosphoS448 mTOR and need to be stimulated with pravastin 2uM and insulin 1ugl/ml (1hr) to have a significant level of the phosphorylated protein. Therefore I strongly suggest that the customer runs the recommended positive control of serum starved 293T cell lysate (preferably nuclear lysate) and to also do a nuclear extraction step to concentrate the protein in her samples. Please ask her to try a 1:500 dilution of the antibody and make sure the secondary antibody is not damaged. I also noticed the customer blocked with two blocking agents (BSA and milk), this can cause problems too so ask her to do a 1hr 5%BSA block and use only TBST to dilute the primary and secondary antibodies (milk contains casein which is a phospho protein so causes interference with phospho-specific antibodies). Please let me know if you need any further assistance,

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