Recombinant
RabMAb

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

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, IPmore 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.

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

  • 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.

  • 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.

  • 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

  • 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 secondary antibody (HRP) (ab131366), was used as secondary antibody 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

ab206340 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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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
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Submitted Sep 07 2017

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