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Anti-PKN1 antibody [EPR18808] (ab195264)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18808] to PKN1
  • Suitable for: IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

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

  • Description

    Rabbit monoclonal [EPR18808] to PKN1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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

     

     

    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.

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

Properties

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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