Anti-PI 3 Kinase p110 delta antibody [EPR386] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PI 3 Kinase p110 delta antibody [EPR386] - BSA and Azide free (AB236049)

Immunocytochemistry/ Immunofluorescence analysis of RAW264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling PI 3 Kinase p110 delta with Purified ab109006 at 1 : 150 dilution (9.8 µg/ml). Cells were fixed in 100% Methanol and permeabilized with none. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^®594) 1 : 200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor^®488, ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109006).

• WB

Lab

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] - BSA and Azide free (AB236049)

This data was developed using ab109006, the same antibody clone in a different buffer formulation.

Western blot : Anti-PI 3 Kinase p110 delta antibody [EPR386] ab109006 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 120 kDa in Wild-type PC-3 cell lysates with no signal observed at this size in PIK3CD knockout PC-3 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (<a href='/ko/products/primary-antibodies/pi-3-kinase-p110-delta-antibody-epr386-ab109006'>ab109006</a>) at 1/1000 dilution

Lane 1:

Wild-type PC-3 at 20 µg

Lane 2:

PIK3CD knockout PC-3 at 20 µg

Lane 3:

Jurkat at 20 µg

Lane 4:

K562 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 120 kDa

Observed band size: 120 kDa

false

• WB

Lab

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] - BSA and Azide free (AB236049)

Lane 1 : Wild type HAP1 whole cell lysate (40 μg)
Lane 2 : PIK3CD knockout HAP1 whole cell lysate (40 μg)
Lane 3 : Jurkat whole cell lysate (40 μg)
Lane 4 : K562 whole cell lysate (40 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab109006 observed at 120 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109006 was shown to specifically react with PIK3CD in wild-type HAP1 cells. No band was observed when PIK3CD knockout samples were used. Wild-type and PIK3CD knockout samples were subjected to SDS-PAGE. ab109006 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 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 1/10,000 dilution for 1hr at room temperature before imaging.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109006).

All lanes:

Western blot - Anti-PI 3 Kinase p110 delta antibody [EPR386] (<a href='/ko/products/primary-antibodies/pi-3-kinase-p110-delta-antibody-epr386-ab109006'>ab109006</a>)

Predicted band size: 119 kDa

Observed band size: 120 kDa

false