Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PRAS40 antibody [EPR6263(2)] (ab151719)

Overview

  • Product name

    Anti-PRAS40 antibody [EPR6263(2)]
    See all PRAS40 primary antibodies
  • Description

    Rabbit monoclonal [EPR6263(2)] to PRAS40
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, IP, Flow Cytmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human PRAS40 aa 150-250. The exact sequence is proprietary.
    Database link: Q96B36

  • Positive control

    • 293T, SHSY5Y, HeLa and NIH 3T3 cell lysates; Human liver tissue; 293T cells.
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab151719 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 27 kDa.
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP 1/10 - 1/100.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    • Function

      Subunit of mTORC1, 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 a 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-389', which then promotes protein synthesis by phosphorylating PDCD4 and targeting it for degradation. Within mTORC1, AKT1S1 negatively regulates mTOR activity in a manner that is dependent on its phosphorylation state and binding to 14-3-3 proteins. Inhibits RHEB-GTP-dependent mTORC1 activation. Substrate for AKT1 phosphorylation, but can also be activated by AKT1-independent mechanisms. May also play a role in nerve growth factor-mediated neuroprotection.
    • Tissue specificity

      Widely expressed with highest levels of expression in liver and heart. Expressed at higher levels in cancer cell lines (e.g. A549 and HeLa) than in normal cell lines (e.g. HEK293).
    • Post-translational
      modifications

      Phosphorylated by AKT1. Phosphorylation relieves inhibitory function on mTORC1.
    • Cellular localization

      Cytoplasm > cytosol. Found in the cytosolic fraction of the brain.
    • Information by UniProt
    • Database links

    • Alternative names

      • 40 kDa proline rich AKT substrate antibody
      • 40 kDa proline-rich AKT substrate antibody
      • AKT1 S1 antibody
      • AKT1 substrate 1 (proline rich) antibody
      • AKT1 substrate 1 antibody
      • AKT1S 1 antibody
      • AKT1S1 antibody
      • AKTS1_HUMAN antibody
      • Lobe antibody
      • MGC2865 antibody
      • PRAS 40 antibody
      • PRAS antibody
      • PRAS40 antibody
      • Proline rich akt substrate antibody
      • Proline rich Akt substrate 40 kDa antibody
      • Proline-rich AKT1 substrate 1 antibody
      see all

    Images

    • All lanes : Anti-PRAS40 antibody [EPR6263(2)] (ab151719) at 1/1000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : AKT1S1 knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : MCF-7 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 27 kDa
      Observed band size: 27 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab151719 observed at 27 kDa. Red - loading control, ab130007, observed at 124 kDa.

      ab151719 was shown to recognize PRAS40 in wild-type HAP1 cells as signal was lost at the expected MW in AKT1S1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and AKT1S1 knockout samples were subjected to SDS-PAGE. Ab151719 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling PRAS40 with ab151719 at 1/50 dilution.
    • All lanes : Anti-PRAS40 antibody [EPR6263(2)] (ab151719) at 1/1000 dilution

      Lane 1 : 293T cell lysate
      Lane 2 : SHSY5Y cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : NIH 3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 27 kDa

    • Flow cytometric analysis of permeabilized 293T cells labeling PRAS40 with ab151719 at 1/10 dilution (red), compared to a nonspecific control antibody (green).

    References

    ab151719 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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