Recombinant
RabMAb

Recombinant Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)

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Overview

  • Product name

    Anti-PDPK1 (phospho S393) antibody [EPR19818]
    See all PDPK1 primary antibodies

  • Description

    Rabbit monoclonal [EPR19818] to PDPK1 (phospho S393)

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IP, Dot blotmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human PDPK1 aa 350-450 (phospho S393). The exact sequence is proprietary.
    Database link: O15530

  • Positive control

    • WB: HEK-293T whole cell lysate transfected with PDPK1 (WT). IP: HEK-293T whole cell lysate transfected with PDPK1 (WT).

  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab206340 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. Detects a band of approximately 63 kDa (predicted molecular weight: 63 kDa).
IP 1/40.
Dot blot 1/1000.

Target

  • Function

    Phosphorylates and activates not only PKB/AKT, but also PKA, PKC-zeta, RPS6KA1 and RPS6KB1. May play a general role in signaling processes and in development (By similarity). Isoform 3 is catalytically inactive.

  • Tissue specificity

    Appears to be expressed ubiquitously.

  • Sequence similarities

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PDK1 subfamily.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.

  • Post-translational
    modifications

    Phosphorylated on tyrosine and serine/threonine. Phosphorylation on Ser-241 in the activation loop is required for full activity. PDK1 itself can autophosphorylate Ser-241, leading to its own activation.

  • Cellular localization

    Cytoplasm. Membrane. Membrane-associated after cell stimulation leading to its translocation. Tyrosine phosphorylation seems to occur only at the plasma membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • 3 phosphoinositide dependent protein kinase 1 antibody
      • 3-phosphoinositide-dependent protein kinase 1 antibody
      • hPDK 1 antibody
      • hPDK1 antibody
      • MGC20087 antibody
      • MGC35290 antibody
      • OTTHUMP00000159109 antibody
      • OTTHUMP00000159110 antibody
      • OTTHUMP00000174525 antibody
      • PDK1 antibody
      • Pdpk1 antibody
      • PDPK1_HUMAN antibody
      • PDPK2 antibody
      • PDPK2P antibody
      • PkB kinase antibody
      • PkB kinase like gene 1 antibody
      • PkB like 1 antibody
      • PRO0461 antibody
      • Protein kinase antibody
      see all

    Images

    • Western blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      Western blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      All lanes : Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340) at 1/5000 dilution

      Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate transfected with PDPK1 S393A mutant
      Lane 2 : HEK-293T whole cell lysate transfected with PDPK1 (WT)
      Lane 3 : HEK-293T whole cell lysate transfected with PDPK1 (WT), then treated with alkaline phosphatase for 1 hour

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 63 kDa
      Observed band size: 60,63 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      These plasmids were kindly provided by our collaborator Dr. Yi Rao, Peking University.

      Based on sequence alignment, the antigen can recognize 5 isoforms, the predicted MW are 63kDa, 58 kDa, 60 kDa, 48 kDa  and 51kDa, respectively.

    • Dot Blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      Dot Blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)

      Dot blot analysis of PDPK1 (phospho S393) labeled with ab206340 at 1/1000 dilution.

      Lane 1: PDPK1 (phospho S393) peptide(aa391-401)

      Lane 2: PDPK1 (phospho S393) peptide(aa386-396)

      Lane 3: PDPK1 non-phospho peptide(aa386-401)

      Lane 4: PDPK1 (phospho S396) peptide (aa392-401)

      Lane 5: PDPK1 (phospho S396) peptide(aa393-402)

      Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated (ab97051) at 1/20000 dilution was used as secondary antibody.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Western blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      Western blot - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      All lanes : Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340) at 1/1000 dilution

      Lane 1 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate transfected with PDPK1 WT
      Lane 2 : HEK-293T whole cell lysate transfected with PDPK1 (WT), then treated with alkaline phosphatase for 1 hour
      Lane 3 : HEK-293T whole cell lysate transfected with PDPK1 S393A mutant

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/2000 dilution

      Predicted band size: 63 kDa
      Observed band size: 60,63 kDa why is the actual band size different from the predicted?


      Exposure time: 5 seconds


      Blocking/Dilution buffer: 5% BSA/TBST.

      This data was kindly provided by our collaborator Dr. Yi Rao, Peking University

    • Immunoprecipitation - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)
      Immunoprecipitation - Anti-PDPK1 (phospho S393) antibody [EPR19818] (ab206340)

      PDPK1 (phospho S393) was immunoprecipitated from 0.35 mg of HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate, transfected with PDPK1 (WT), with ab206340 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab206340 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HEK-293T whole cell lysate transfected with PDPK1 (WT), 10 μg (Input).

      Lane 2: ab206340 IP in HEK-293T whole cell lysate transfected with PDPK1 (WT).

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206340 in HEK-293T whole cell lysate transfected with PDPK1 (WT).

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3 minutes.

      The plasmids were kindly provided by our collaborator Dr. Yi Rao, Peking University.

    References

    This product has been referenced in:

    • Wang C  et al. TCRP1 promotes NIH/3T3 cell transformation by over-activating PDK1 and AKT1. Oncogenesis 6:e323 (2017). WB ; Human . Read more (PubMed: 28436990) »
    See 1 Publication for this product

    Publishing research using ab206340? Please let us know so that we can cite the reference in this datasheet.

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    Western blot abreview for Anti-PDPK1 (phospho S393) antibody [EPR19818]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (293t)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    10 µg
    Specification
    293t
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 30°C
    Read More

    Abcam user community

    Verified customer

    Submitted Sep 07 2017

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