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

• IP

Supplier Data

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.

All lanes:

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

Predicted band size: 124 kDa

Observed band size: 110 kDa

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• WB

Lab

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.

All lanes:

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

Predicted band size: 124 kDa

false

• WB

Supplier Data

Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (AB183957)

Blocking/Dilution buffer : 5% NFDM/TBST.

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

All lanes:

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

All lanes:

P0 Rat brain lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 124 kDa

Observed band size: 110 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EPR19693] (AB183957)

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).

All lanes:

Western blot - 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 at 20 µg

Lane 2:

M1 (Mouse myeloblast myeloid leukemia cell line) whole cell lysate at 20 µg

Lane 3:

Mouse hypothalamus lysate at 20 µg

Lane 4:

Mouse fetal brain lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 124 kDa

Observed band size: 110 kDa

false