Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18416] to PI 3 Kinase p85 beta - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, IP
  • Knockout validated
  • Reacts with: Rat, Human

Overview

  • Product name

    Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free
    See all PI 3 Kinase p85 beta primary antibodies

  • Description

    Rabbit monoclonal [EPR18416] to PI 3 Kinase p85 beta - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt

  • Species reactivity

    Reacts with: Rat, Human

  • Immunogen

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

  • Positive control

    • WB: HeLa, Jurkat, HEK293 cell lysates; Human fetal brain and fetal kidney lysates; Rat brain and spleen lysates; PC12 cell lysate. IHC-P: Human colonic adenocarcinoma, Human colon, rat testis tissues. ICC/IF: HeLa and Jurkat cells. IP: Jurkat cell lysate.

  • General notes

    ab231161 is the carrier-free version of ab180967.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

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

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Images

  • Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Western blot - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    All lanes : Anti-PI 3 Kinase p85 beta antibody [EPR18416] (ab180967) at 1/2000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : PIK3R2 knockout HEK-293T cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 82 kDa
    Observed band size: 85 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 containing PBS, BSA, glycerol, and sodium azide (ab180967).

    Lanes 1- 2: Merged signal (red and green). Green - ab180967 observed at 85 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab180967 was shown to react with PI 3 Kinase p85 beta in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266799 (knockout cell lysate ab257586) was used. Wild-type HEK-293T and PIK3R2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. ab180967 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Nucleus and cytoplasm staining on spermatogenic cell of rat testis was observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180967).

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling PI 3 Kinase p85 beta with ab180967 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and cytoplasmic staining on HeLa cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab180967 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) 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 (ab180967).

  • Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling PI 3 Kinase p85 beta with ab180967 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Confocal image showing nuclear and weakly cytoplasmic staining on Jurkat cell line.

    The nuclear counter stain is DAPI (blue).

    Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
    The negative controls are as follows:
    1. ab180967 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) 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 (ab180967).

  • Immunoprecipitation - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunoprecipitation - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    PI 3 Kinase p85 beta was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab180967 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab180967 at 1/10000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: HeLa whole cell lysate10 µg (Input).

    Lane 2: ab180967 IP in HeLa whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab180967 in HeLa whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 10 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180967).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    This IHC data was generated using the same anti-PI 3 Kinase p85 beta antibody clone, EPR18416, in a different buffer formulation (cat# ab180967).

    Immunohistochemical analysis of paraffin-embedded Human colonic adenocarcinoma tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Nucleus and cytoplasm staining on tumor cells of colonic adenocarcinoma was observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

    This IHC data was generated using the same anti-PI 3 Kinase p85 beta antibody clone, EPR18416, in a different buffer formulation (cat# ab180967).

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling PI 3 Kinase p85 beta using ab180967 at 1/100 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) was used as secondary at 1/500 dilution. Cytoplasm staining on epithelial cells of Human colon was observed. Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)
    Anti-PI 3 Kinase p85 beta antibody [EPR18416] - BSA and Azide free (ab231161)

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