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  1. Link

    pras40-antibody-epr6402-ab133584.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Other
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-PRAS40 antibody [EPR6402] (ab133584)

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  • SDS
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Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
  • Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
  • OI-RD Scanning - Anti-PRAS40 antibody [EPR6402] (ab133584)
  • Anti-PRAS40 antibody [EPR6402] (ab133584)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6402] to PRAS40
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-PRAS40 antibody [EPR6402]
    See all PRAS40 primary antibodies
  • Description

    Rabbit monoclonal [EPR6402] to PRAS40
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human PRAS40 aa 150-250 (internal sequence). The exact sequence is proprietary.

  • Positive control

    • MCF7, L6, C6, RAW 264.7, PC12, NIH 3T3, 293T and HeLa whole cell lysate (ab150035)
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Dissociation constant (KD)

    KD = 1.43 x 10 -11 M
    Learn more about KD
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR6402
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Other
    • Neuroscience
    • Neurotransmission
    • Intracellular Signaling
    • Kinases
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases

Associated products

  • Alternative Versions

    • Anti-PRAS40 antibody [EPR6402] - BSA and Azide free (ab248564)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • RAW 264.7 whole cell lysate (ab7187)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab133584 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. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa).
Notes
WB
1/1000 - 1/10000. Detects a band of approximately 40 kDa (predicted molecular weight: 27 kDa).
Application notes
Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

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.
  • Target information above from: UniProt accession Q96B36 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 84335 Human
    • Entrez Gene: 67605 Mouse
    • Entrez Gene: 292887 Rat
    • Omim: 610221 Human
    • SwissProt: Q96B36 Human
    • SwissProt: Q9D1F4 Mouse
    • Unigene: 515542 Human
    • Unigene: 148007 Mouse
    • 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

    • Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
      Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
      All lanes : Anti-PRAS40 antibody [EPR6402] (ab133584) 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: 40 kDa why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab133584 observed at 40 kDa. Red - loading control, ab130007, observed at 125 kDa.

      ab133584 was shown to specifically react with PRAS40 in wild-type HAP1 cells as signal was lost in AKT1S1 (PRAS40) knockout cells. Wild-type and AKT1S1 (PRAS40) knockout samples were subjected to SDS-PAGE. Ab133584 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.

    • Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
      Western blot - Anti-PRAS40 antibody [EPR6402] (ab133584)
      All lanes : Anti-PRAS40 antibody [EPR6402] (ab133584) at 1/1000 dilution

      Lane 1 : MCF7 cell lysate
      Lane 2 : L6 cell lysate
      Lane 3 : C6 cell lysate
      Lane 4 : RAW 264.7 cell lysate
      Lane 5 : PC12 cell lysate
      Lane 6 : NIH 3T3 cell lysate
      Lane 7 : 293T cell lysate
      Lane 8 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 27 kDa

    • OI-RD Scanning - Anti-PRAS40 antibody [EPR6402] (ab133584)
      OI-RD Scanning - Anti-PRAS40 antibody [EPR6402] (ab133584)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-PRAS40 antibody [EPR6402] (ab133584)
      Anti-PRAS40 antibody [EPR6402] (ab133584)

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (0)

    Publishing research using ab133584? Please let us know so that we can cite the reference in this datasheet.

    ab133584 has not yet been referenced specifically in any publications.

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