Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-PKN1 antibody [EPR18808] (ab195264)

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Overview

  • Product name

    Anti-PKN1 antibody [EPR18808]
    See all PKN1 primary antibodies

  • Description

    Rabbit monoclonal [EPR18808] to PKN1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human PKN1 aa 300-500. The exact sequence is proprietary.
    Database link: Q16512

  • Positive control

    • WB: Jurkat, LNCaP, MDA-MB-231, K562, HEK-293, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Rat brain and spleen lysates. IP: Jurkat whole cell lysate.

  • 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 ab195264 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
IP 1/40.
WB 1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 104 kDa).

Target

  • Function

    PKC-related serine/threonine-protein kinase involved in various processes such as regulation of the intermediate filaments of the actin cytoskeleton, cell migration, tumor cell invasion and transcription regulation. Regulates the cytoskeletal network by phosphorylating proteins such as VIM and neurofilament proteins NEFH, NEFL and NEFM, leading to inhibit their polymerization. Phosphorylates 'Ser-575', 'Ser-637' and 'Ser-669' of MAPT/Tau, lowering its ability to bind to microtubules, resulting in disruption of tubulin assembly. Acts as a key coactivator of androgen receptor (ANDR)-dependent transcription, by being recruited to ANDR target genes and specifically mediating phosphorylation of 'Thr-11' of histone H3 (H3T11ph), a specific tag for epigenetic transcriptional activation that promotes demethylation of histone H3 'Lys-9' (H3K9me) by KDM4C/JMJD2C. Phosphorylates HDAC5, HDAC7 and HDAC9, leading to impair their import in the nucleus. Phosphorylates 'Thr-38' of PPP1R14A, 'Ser-159', 'Ser-163' and 'Ser-170' of MARCKS, and GFAP. Able to phosphorylate RPS6 in vitro.

  • Tissue specificity

    Found ubiquitously. Expressed in heart, brain, placenta, lung, skeletal muscle, kidney and pancreas. Expressed in numerous tumor cell lines, especially in breast tumor cells.

  • Sequence similarities

    Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 C2 domain.
    Contains 1 protein kinase domain.
    Contains 3 REM (Hr1) repeats.

  • Domain

    The C1 domain does not bind the diacylglycerol (DAG).

  • Post-translational
    modifications

    Autophosphorylated; preferably on serine. Phosphorylated during mitosis.
    Activated by limited proteolysis with trypsin.

  • Cellular localization

    Cytoplasm. Nucleus. Endosome. Cell membrane. Cleavage furrow. Midbody. Associates with chromatin in a ligand-dependent manner. Localization to endosomes is mediated via its interaction with RHOB. Association to the cell membrane is dependent on Ser-374 phosphorylation. Accumulates during telophase at the cleavage furrow and finally concentrates around the midbody in cytokinesis.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • DBK antibody
    • PAK 1 antibody
    • PAK-1 antibody
    • PAK1 antibody
    • PKC1 antibody
    • PKN ALPHA antibody
    • PKN antibody
    • Pkn1 antibody
    • PKN1_HUMAN antibody
    • PRK1 antibody
    • PRKCL1 antibody
    • Protease activated kinase 1 antibody
    • Protease-activated kinase 1 antibody
    • Protein kinase C like 1 antibody
    • Protein kinase C like PKN antibody
    • Protein kinase C related kinase 1 antibody
    • Protein kinase C-like 1 antibody
    • Protein kinase C-like PKN antibody
    • Protein kinase N1 antibody
    • Protein kinase PKN alpha antibody
    • Protein kinase PKN-alpha antibody
    • Protein-kinase C-related kinase 1 antibody
    • Serine threonine kinase N antibody
    • Serine threonine protein kinase N antibody
    • Serine-threonine protein kinase N antibody
    • Serine/threonine protein kinase N1 antibody
    • Serine/threonine-protein kinase N1 antibody
    see all

Images

  • Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)

    Lane 1: Wild-type HAP1 cell lysate (40 µg)
    Lane 2: PKN1 knockout HAP1 cell lysate (40 µg)
    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: K562 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab195264 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab195264 was shown to specifically react with PKN1 when PKN1 knockout samples were used. Wild-type and PKN1 knockout samples were subjected to SDS-PAGE. Ab195264 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 dilution respectively and incubated overnight at 4C. 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.

  • Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    All lanes : Anti-PKN1 antibody [EPR18808] (ab195264) at 1/1000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : LNCaP (Human prostate cancer cell line) whole cell lysate
    Lane 3 : MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 4 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
    Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 6 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size: 104 kDa
    Observed band size: 120 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunoprecipitation - Anti-PKN1 antibody [EPR18808] (ab195264)
    Immunoprecipitation - Anti-PKN1 antibody [EPR18808] (ab195264)

    PKN1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab195264 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab195264 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Jurkat whole cell lysate, 10µg (Input).

    Lane 2: ab195264 IP in Jurkat whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal-Isotype Control (ab172730) instead of ab195264 in Jurkat whole cell lysate.

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

    Exposure time: 3 minutes.

  • Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    All lanes : Anti-PKN1 antibody [EPR18808] (ab195264) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    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: 104 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    Western blot - Anti-PKN1 antibody [EPR18808] (ab195264)
    All lanes : Anti-PKN1 antibody [EPR18808] (ab195264) at 1/1000 dilution

    Lane 1 : Rat brain lysate
    Lane 2 : Rat spleen lysate

    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: 104 kDa
    Observed band size: 120 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes

References

This product has been referenced in:

  • Chang J  et al. Exome-wide analysis identifies three low-frequency missense variants associated with pancreatic cancer risk in Chinese populations. Nat Commun 9:3688 (2018). Read more (PubMed: 30206226) »
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