Anti-AP4E1 抗体 [EPR28049-87] - BSA and Azide free

• WB

Lab

Western blot - Anti-AP4E1 antibody [EPR28049-87] - BSA and Azide free (AB318190)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab318189).

Western blot : Anti-AP4E1 antibody [EPR28049-87] ab318189 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 127 kDa in Wild-type SH-SY5Y cell lysates with no signal observed at this size in AP4E1 knockout SH-SY5Y cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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-AP4E1 antibody [EPR28049-87] (<a href='/products/primary-antibodies/ap4e1-antibody-epr28049-87-ab318189'>ab318189</a>) at 1/1000 dilution

Lane 1:

Wild-type SH-SY5Y cell lysates at 20 µg

Lane 2:

AP4E1 knockout SH-SY5Y cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

PC-3 cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 127 kDa

Observed band size: 127 kDa

false

• WB

Supplier Data

Western blot - Anti-AP4E1 antibody [EPR28049-87] - BSA and Azide free (AB318190)

This data was developed using ab318189, the same antibody clone in a different buffer formulation.

The bands beneath the target band (Lane 2, 25~75 kDa) are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 103 seconds; Lane 2 : 26 seconds

All lanes:

Western blot - Anti-AP4E1 antibody [EPR28049-87] (<a href='/products/primary-antibodies/ap4e1-antibody-epr28049-87-ab318189'>ab318189</a>) at 1/1000 dilution

Lane 1:

Human brainstem tissue lysate at 20 µg

Lane 2:

Human placenta tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 127 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-AP4E1 antibody [EPR28049-87] - BSA and Azide free (AB318190)

This data was developed using ab318189, the same antibody clone in a different buffer formulation.

The bands beneath the target band (Lane 1-3, 25~37 kDa) are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1, 4 : 180 seconds; Lane 2, 3, 5 : 136 seconds

All lanes:

Western blot - Anti-AP4E1 antibody [EPR28049-87] (<a href='/products/primary-antibodies/ap4e1-antibody-epr28049-87-ab318189'>ab318189</a>) at 1/1000 dilution

Lane 1:

HaCaT (human skin keratinocyte) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole fresh cell lysate at 20 µg

Lane 5:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole fresh cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 127 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-AP4E1 antibody [EPR28049-87] - BSA and Azide free (AB318190)

This data was developed using ab318189, the same antibody clone in a different buffer formulation.

The identity of the lower MW bands at approximately 60kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-AP4E1 antibody [EPR28049-87] (<a href='/products/primary-antibodies/ap4e1-antibody-epr28049-87-ab318189'>ab318189</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 40 µg

Lane 2:

HeLa transfected with siRNA specifically targeting AP4E1 whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 127 kDa,36 kDa

false

Exposure time: 136s