Anti-ESE1 antibody [EPESER1] - BSA and Azide free

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-ESE1 antibody [EPESER1] - BSA and Azide free (AB248579)

This data was developed using ab133621, the same antibody clone in a different buffer formulation.

Overlay histogram showing A431 cells stained with ab133621 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133621, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

• WB

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Western blot - Anti-ESE1 antibody [EPESER1] - BSA and Azide free (AB248579)

This data was developed using ab133621, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-ESE1 antibody [EPESER1] (<a href='/en-us/products/primary-antibodies/ese1-antibody-epeser1-ab133621'>ab133621</a>) at 1/1000 dilution

Lane 1:

Human uterus cell lysate at 10 µg

Lane 2:

Fetal lung cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

C6 cell lysate at 10 µg

Lane 5:

RAW 264.7 cell lysate at 10 µg

Lane 6:

PC12 cell lysate at 10 µg

Lane 7:

NIH3T3 cell lysate at 10 µg

Lane 8:

SKBR-3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 41 kDa

Observed band size: 47 kDa

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• WB

Lab

Western blot - Anti-ESE1 antibody [EPESER1] - BSA and Azide free (AB248579)

This data was developed using ab133621, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPESER1] to ESE1 ab133621 staining at 1/1000 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 41 kDa in Wild-type A549 cell lysates with no signal observed at this size in ELF3 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-ESE1 antibody [EPESER1] (<a href='/en-us/products/primary-antibodies/ese1-antibody-epeser1-ab133621'>ab133621</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

ELF3 knockout A549 at 20 µg

Lane 3:

A431 at 20 µg

Secondary

All lanes:

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

Predicted band size: 41 kDa

Observed band size: 41 kDa

false