Anti-PDPK1 antibody [EPR245] (ab109253)

Overview

  • Product nameAnti-PDPK1 antibody [EPR245]
    See all PDPK1 primary antibodies
  • Description
    Rabbit monoclonal [EPR245] to PDPK1
  • Tested applicationsICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PDPK1 aa 1-100 (N terminal).
    Database link: O15530

  • Positive control
    • 293T and PC12 cell lysates; Human breast carcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    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.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Alternative versions available:

    Anti-PDPK1 antibody (Alexa Fluor® 647) [EPR245] (ab208125)

    Anti-PDPK1 antibody (Alexa Fluor® 488) [EPR245] (ab208124)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab109253 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
ICC/IF 1/100 - 1/250.
WB 1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Target

  • FunctionPhosphorylates 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 specificityAppears to be expressed ubiquitously.
  • Sequence similaritiesBelongs 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 localizationCytoplasm. 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
    • PkB kinase antibody
    • PkB kinase like gene 1 antibody
    • PkB like 1 antibody
    • PRO0461 antibody
    • Protein kinase antibody
    see all

Anti-PDPK1 antibody [EPR245] images

  • Anti-PDPK1 antibody [EPR245] (ab109253) at 1/5000 dilution (purified) + mouse brain at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 63 kDa
    Observed band size : 58,68 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Anti-PDPK1 antibody [EPR245] (ab109253) at 1/4000 dilution (purified) + Jurkat cell lysate at 10 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 63 kDa
    Observed band size : 58,68 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Anti-PDPK1 antibody [EPR245] (ab109253) at 1/1000 dilution (purified) + HEK293 cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 63 kDa
    Observed band size : 58,68 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of HEK293 cells with purified ab109253 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab109253 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human kidney with purified ab109253 at a working dilution of 1/500. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • All lanes : Anti-PDPK1 antibody [EPR245] (ab109253) at 1/1000 dilution (unpurified)

    Lane 1 : 293T cell lysate
    Lane 2 : PC12 cell lysate

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 63 kDa
  • Unpurified ab109253, at a 1/500 dilution, staining PDPK1 in Human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

References for Anti-PDPK1 antibody [EPR245] (ab109253)

ab109253 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (head and neck cancer)
Gel Running Conditions Reduced Denaturing (10)
Loading amount 20 µg
Treatment DCA 20mM for 24hs
Specification head and neck cancer
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 02 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (head and neck cancer)
Gel Running Conditions Non-reduced Non-Denaturing (Native) (gel 10%)
Loading amount 30 µg
Treatment DCA 20mM for 24hs
Specification head and neck cancer
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Miss. jin young park

Verified customer

Submitted May 22 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (head and neck cancer)
Permeabilization Yes - triton x 100
Specification head and neck cancer
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Apr 28 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"