Anti-INPP4A antibody [EP3425(2)] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-INPP4A antibody [EP3425(2)] - BSA and Azide free (AB247910)

This data was developed using ab109622, the same antibody clone in a different buffer formulation.

ab109622 at 1/100 dilution staining INPP4A in paraffin-embedded Human brain tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-INPP4A antibody [EP3425(2)] - BSA and Azide free (AB247910)

This data was developed using ab109622, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : INPP4A knockout HAP1 cell lysate (20 μg)

Lane 3 : Jurkat cell lysate (20 μg)

Lane 4 : Human fetal brain tissue lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109622 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109622 was shown to specifically react with INPP4A when INPP4A knockout samples were used, along with additional cross-reactive bands. Wild-type and INPP4A knockout samples were subjected to SDS-PAGE. ab109622 and ab8245 (loading control to GAPDH) were diluted at 1/500 and 1/10,000 dilution respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) andGoat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-INPP4A antibody [EP3425(2)] (<a href='/en-us/products/primary-antibodies/inpp4a-antibody-ep34252-ab109622'>ab109622</a>)

Predicted band size: 110 kDa

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• WB

Lab

Western blot - Anti-INPP4A antibody [EP3425(2)] - BSA and Azide free (AB247910)

This data was developed using ab109622, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EP3425(2)] to INPP4A ab109622 staining at 1/500 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 110 kDa in Wild-type A549 cell lysates with no signal observed at this size in INPP4A knockout A549 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-INPP4A antibody [EP3425(2)] (<a href='/en-us/products/primary-antibodies/inpp4a-antibody-ep34252-ab109622'>ab109622</a>) at 1/500 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

INPP4A knockout A549 at 20 µg

Lane 3:

HL-60 at 20 µg

Lane 4:

MCF7 at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 110 kDa

Observed band size: 110 kDa,50 kDa

false

• WB

Unknown

Western blot - Anti-INPP4A antibody [EP3425(2)] - BSA and Azide free (AB247910)

This data was developed using ab109622, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-INPP4A antibody [EP3425(2)] (<a href='/en-us/products/primary-antibodies/inpp4a-antibody-ep34252-ab109622'>ab109622</a>) at 1/1000 dilution

Lane 1:

Fetal brain lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Secondary

All lanes:

HRP-labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 110 kDa

Observed band size: 104 kDa

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