Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18702] to PI 3 Kinase p85 alpha - BSA and Azide free
  • Suitable for: Flow Cyt, ICC/IF, IP, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free
    See all PI 3 Kinase p85 alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR18702] to PI 3 Kinase p85 alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    ICC/IF
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human PI3K p85 alpha full length recombinant protein; Human fetal liver, fetal heart and fetal kidney lysates; HeLa, HepG2, MCF7, Raji, Jurkat, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, heart, kidney and spleen lysates. ICC/IF: HepG2 and NIH/3T3 cells. Flow Cyt: NIH/3T3 cells; IP: MCF7 whole cell lysate.

  • General notes

    Ab223792 is the carrier-free version of ab191606. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab223792 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    This WB data was generated using the same anti-PI 3 Kinase p85 alpha antibody clone [EPR18702] in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (cat# ab191606).

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: PIK3R1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Jurkat whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab191606 was shown to specifically react with PIK3R1 when PIK3R1 knockout samples were used. Wild-type and PIK3R1 knockout samples were subjected to SDS-PAGE. Ab191606 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 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.

  • Flow Cytometry - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Flow Cytometry - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    Overlay histogram showing HepG2 cells fixed in 4% PFA and stained with ab191606 at a dilution of 1/80 (red line). The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG (ab172730) was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191606).

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PI3K p85 with ab191606 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291)  at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120)  at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab191606 at 1/500 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291  at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191606).

  • Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    All lanes : Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (ab191606) 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: 84 kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab191606).

    Lanes 1- 2: Merged signal (red and green). Green - ab191606 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab191606 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. ab191606 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 100% Methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PI3K p85 with ab191606 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at  1/1000 dilution and Goat Anti-Mouse IgG  (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab191606 at 1/500 dilution followed by ab150120  at 1/1000 dilution.
    -ve control 2: ab7291 at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191606).

  • Flow Cytometry - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Flow Cytometry - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PI3K p85 with ab191606 at 1/150 dilution (red) compared with a Rabbit IgG,monoclonal -  Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191606).

  • Immunoprecipitation - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Immunoprecipitation - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

    PI3K p85 was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell lysate with ab191606 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191606 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: MCF7 whole cell lysate, 10µg (Input).

    Lane 2: ab191606 IP in MCF7 whole cell lysate.

    Lane 3: Rabbit IgG, monoclonal - Isotype Control (ab172730) instead of ab191606 in MCF7 whole cell lysate.

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

    Exposure time: 5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191606).

  • Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)
    Anti-PI 3 Kinase p85 alpha antibody [EPR18702] - BSA and Azide free (ab223792)

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