Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free

5 product images

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  Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)

  This WB data was generated using the same anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody clone, EP383Y, in a different buffer formulation (cat# Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776).
  

  Lane 1: Wild-type HAP1 cell lysate (20 μg)
  Lane 2: PI 3 Kinase catalytic subunit alpha/PIK3CA knockout HAP1 cell lysate (20 μg)
  Lane 3: Jurkat cell lysate (20 μg)
  Lane 4: MCF7 cell lysate (20 μg)
  Lanes 1 - 4: Merged signal (red and green). Green - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 observed at 120 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
  
  Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 was shown to recognize PI 3 Kinase catalytic subunit alpha when PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were used, along with additional cross-reactive bands. Wild-type and PI 3 Kinase catalytic subunit alpha/PIK3CA knockout samples were subjected to SDS-PAGE. Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776)

  Predicted band size: 124 kDa

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  Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)

  Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 (purified) at a dilution of 1/20 immunoprecipitating PI 3 Kinase catalytic subunit alpha/PIK3CA in Jurkat whole cell lysate.
  

  Lane 1 (input): Jurkat whole cell lysate (10μg)

  Lane 2 (+): Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 + Jurkat whole cell lysate.

  Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 in Jurkat whole cell lysate.

  For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.

  Blocking buffer and concentration: 5% NFDM/TBST.

  Diluting buffer and concentration: 5% NFDM /TBST.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776).

  All lanes: Immunoprecipitation - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776)

  Predicted band size: 124 kDa

  Observed band size: 110 kDa

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  Flow Cytometry (Intracellular) - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)

  Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 staining PI 3 Kinase catalytic subunit alpha/PIK3CA in the human cell line Jurkat (human acute T cell leukemia) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permiabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.
  

  Isoytype control: Rabbit monoclonal IgG (Black)

  Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776).

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  Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)

  Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with unpurified Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 at a dilution of 1/100.

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776).

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  Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] - BSA and Azide free (ab185927)

  This ICC/IF data was generated using the same anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody clone, EP383Y, in a different buffer formulation (cat# Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776).
  

  Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling PI 3 Kinase catalytic subunit alpha/PIK3CA with purified Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] ab40776 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

  Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

  Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).