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Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19693] to PI 3 Kinase catalytic subunit alpha/PIK3CA
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693]
    See all PI 3 Kinase catalytic subunit alpha/PIK3CA primary antibodies

  • Description

    Rabbit monoclonal [EPR19693] to PI 3 Kinase catalytic subunit alpha/PIK3CA

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IP
    Mouse
    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: F9 and M1 whole cell lysates; Mouse hypothalamus and fetal brain lysates; P0 Rat brain lysate. IP: Mouse hypothalamus 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-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: PI 3 Kinase catalytic subunit alpha/PIK3CA knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab183957 observed at 125 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab183957 was shown to recognize PI 3 Kinase catalytic subunit alpha/PIK3CA in wild type cells as signal was lost at the expected MW in PI 3 Kinase catalytic subunit alpha/PIK3CA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were subjected to SDS-PAGE. ab183957 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)

    PI 3 Kinase catalytic subunit alpha/PIK3CA was immunoprecipitated from 0.35 mg of mouse hypothalamus lysate with ab183957 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab183957 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

    Lane 1: Mouse hypothalamus lysate, 10µg (Input).
    Lane 2: ab183957 IP in mouse hypothalamus lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab183957 in mouse hypothalamus lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    All lanes : Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957) at 1/1000 dilution

    Lane 1 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
    Lane 2 : M1 (Mouse myeloblast myeloid leukemia cell line) whole cell lysate
    Lane 3 : Mouse hypothalamus lysate
    Lane 4 : Mouse fetal brain lysate

    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: 124 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1: 3 minutes; Lane 2 and 3: 5 seconds; Lane 4: 3 seconds.

    The observed molecular weight is consistent with what has been described in the literature (PMID: 19701705).

  • Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957) at 1/1000 dilution + P0 Rat brain lysate at 20 µg

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

    Predicted band size: 124 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed molecular weight is consistent with what has been described in the literature (PMID: 19701705).

  • Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)
    Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (ab183957)

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