Anti-PI 3 Kinase p85 alpha antibody [EP380Y] - BSA and Azide free

• WB

Unknown

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] - BSA and Azide free (AB182617)

This data was developed using ab40755, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-ep380y-ab40755'>ab40755</a>) at 1/2000 dilution

Lane 1:

A431 cell lysate at 10 µg

Lane 2:

MCF-7 cell lysate at 10 µg

Lane 3:

Raji cell lysate at 10 µg

Predicted band size: 83 kDa

Observed band size: 85 kDa

false

• WB

Lab

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] - BSA and Azide free (AB182617)

This data was developed using ab40755, the same antibody clone in a different buffer formulation.

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 - ab40755 observed at 90 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab40755 was shown to recognize PI 3 Kinase p85 alpha in wild-type HAP1 cells along with additional cross-reactive bands. No Band was observed when PI 3 Kinase p85 alpha knockout samples were examined. Wild-type and PI 3 Kinase p85 alpha knockout samples were subjected to SDS-PAGE. ab40755 and ab181602 (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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-ep380y-ab40755'>ab40755</a>)

Predicted band size: 83 kDa

false

• WB

Lab

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] - BSA and Azide free (AB182617)

This data was developed using ab182617, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

Compared with ab40755, ab191606 has higher sensitivity. We recommend ab124805 as an alternative for testing western blot.

We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab197896 in western blot.

Lane 1:

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EP380Y] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-ep380y-ab40755'>ab40755</a>) at 1/1000 dilution

Lane 2:

Western blot - Anti-PI 3 Kinase p85 alpha antibody [EPR18702] (<a href='/en-us/products/primary-antibodies/pi-3-kinase-p85-alpha-antibody-epr18702-ab191606'>ab191606</a>) at 1/1000 dilution

All lanes:

Jurkat (human T cell leukemia T lymphocyte) whole cell 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

Observed band size: 85 kDa

false

Exposure time: 40s