Overview

  • Product name

    Anti-PDPK1 antibody [EP569Y]
    See all PDPK1 primary antibodies
  • Description

    Rabbit monoclonal [EP569Y] to PDPK1
  • Host species

    Rabbit
  • Specificity

    This antibody detects PDK1 both when phosphorylated and non-phosphorylated on Tyrosine 373 and 376.

    The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig.

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Synthetic phospho-peptide corresponding to residues surrounding Tyr373 and Tyr376 of human PDPK1.

  • Positive control

    • WB: PC-12, Rat testis, HEK-293, 22Rv1, PIEC, HeLa, K-562, and F9 lysates. IHC-P: human lung carcinoma and human prostate carcinoma tissue.
  • 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.

    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 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab52893 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 64 kDa (predicted molecular weight: 63 kDa).
IHC-P 1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

The mouse, rat and pig recommendation is based on the WB results. We do not guarantee IHC-P for mouse, rat and pig.

See IHC antigen retrieval protocols.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • 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

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: PDPK1 knockout HAP1 cell lysate (20 µg)
      Lane 3: HEK293 cell lysate (20 µg)
      Lane 4: MCF7 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab52893 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab52893 was shown to recognize PDPK1 when PDPK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and PDPK1 knockout samples were subjected to SDS-PAGE. ab52893 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified) + F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

      Predicted band size: 63 kDa
      Observed band size: 52-65 kDa
      why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in PMID: 30171166

    • All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)

      Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
      Lane 2 : K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/5000 dilution

      Predicted band size: 63 kDa
      Observed band size: 52-65 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in PMID: 30171166

    • All lanes : Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution (Purified)

      Lane 1 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
      Lane 2 : Rat testis lysates
      Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates
      Lane 4 : 22Rv1 (Human prostate carcinoma epithelial cell) whole cell lysates
      Lane 5 : PIEC (PIG iliac artery endothelial cell) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 63 kDa
      Observed band size: 52-65 kDa why is the actual band size different from the predicted?



      The molecular weight observed is consistent with what has been described in PMID: 30171166

    • Ab52893 (1:100) staining human PDPK1 in human prostate carcinoma tissue by immunohistochemistry using paraffin embedded tissue.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Anti-PDPK1 antibody [EP569Y] (ab52893) at 1/1000 dilution + 293 cell lysate

      Predicted band size: 63 kDa
      Observed band size: 63 kDa

    References

    This product has been referenced in:

    • Han T  et al. miR-16-2-3p inhibits cell proliferation and migration and induces apoptosis by targeting PDPK1 in maxillary primordium mesenchymal cells. Int J Mol Med 43:1441-1451 (2019). Read more (PubMed: 30664182) »
    • Jin H  et al. Differential contribution of the two waves of cardiac progenitors and their derivatives to aorta and pulmonary artery. Dev Biol 450:82-89 (2019). Read more (PubMed: 30951706) »
    See all 12 Publications for this product

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