Anti-USP13 antibody [EPR4348] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-USP13 antibody [EPR4348] - BSA and Azide free (AB247812)

This data was developed using ab109264, the same antibody clone in a different buffer formulation.

Overlay histogram showing SHSY-5Y cells stained with ab109264 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab109264, 1/9400 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. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

Lab

Western blot - Anti-USP13 antibody [EPR4348] - BSA and Azide free (AB247812)

This data was developed using ab109264, the same antibody clone in a different buffer formulation.

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

Lane 2 : USP13 knockout HAP1 cell lysate (20 μg)

Lane 3 : A375 cell lysate (20 μg)

Lane 4 : SH-SY5Y cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab109264 observed at 100 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109264 was shown to recognize USP13 when USP13 knockout samples were used, along with additional cross-reactive bands. Wild-type and USP13 knockout samples were subjected to SDS-PAGE. ab109264 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/10 000 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-USP13 antibody [EPR4348] (<a href='/en-us/products/primary-antibodies/usp13-antibody-epr4348-ab109264'>ab109264</a>)

Predicted band size: 97 kDa

false

• WB

Unknown

Western blot - Anti-USP13 antibody [EPR4348] - BSA and Azide free (AB247812)

This data was developed using ab109264, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-USP13 antibody [EPR4348] (<a href='/en-us/products/primary-antibodies/usp13-antibody-epr4348-ab109264'>ab109264</a>) at 1/1000 dilution

Lane 1:

SH SY5Y cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

A375 cell lysate at 10 µg

Predicted band size: 97 kDa

Observed band size: 97 kDa

false