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Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PI 3 Kinase p85 alpha antibody [EPR5513]
    See all PI 3 Kinase p85 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR5513] to PI 3 Kinase p85 alpha

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide corresponding to a region within the N terminus of Human PI 3 Kinase p85 alpha (UniProt ID: P27986)

  • Positive control

    • HeLa, Jurkat, Raji, K562, MCF7, C6, Raw 264.7, PC12, NIH3T3 cell lysates

  • 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 p85 alpha antibody [EPR5513] (ab133595)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PI 3 Kinase p85 alpha knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate (20 µg)
    Lane 4: THP1 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab133595 was shown to specifically react with PI 3 Kinase p85 alpha when PI 3 Kinase p85 alpha knockout samples were used. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. ab133595 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 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 h at room temperature before imaging.

  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : PIK3R1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 83 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    Lanes 1- 2: Merged signal (red and green). Green - ab133595 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab133595 was shown to react with PI 3 Kinase p85 alpha in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265116 (knockout cell lysate ab257029) was used. Wild-type HeLa and PIK3R1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133595 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : Raji cell lysate
    Lane 3 : K562 cell lysate
    Lane 4 : MCF7 cell lysate
    Lane 5 : C6 cell lysate
    Lane 6 : Raw 264.7 cell lysate
    Lane 7 : PC12 cell lysate
    Lane 8 : NIH3T3 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled Goat anti Rabbit at 1/2000 dilution

    Predicted band size: 83 kDa

  • OI-RD Scanning - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    OI-RD Scanning - Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)
    Anti-PI 3 Kinase p85 alpha antibody [EPR5513] (ab133595)

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