Anti-BirA antibody [6C4c7] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BirA antibody [6C4c7] - BSA and Azide free (AB269579)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab232732)

ab232732 staining BirA in normal HEK293 cells, BirA transfected HEK293T cells (uninduced) and BirA transfected HEK293T cells induced with DOX. The cells were fixed with 100% MeOH (5min), permeabilized with 0.1%PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab232732 at 5μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Cell samples kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.

• WB

Lab

Western blot - Anti-BirA antibody [6C4c7] - BSA and Azide free (AB269579)

This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 5% milk before ab232732 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1μg/ml concentration and 1/20,000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.

Cell samples kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.

This image was produced using the same antibody clone but in a different formulation ab232732, PBS and sodium azide.

All lanes:

Western blot - Anti-BirA antibody [6C4c7] (<a href='/en-us/products/primary-antibodies/bira-antibody-6c4c7-ab232732'>ab232732</a>) at 1 µg/mL

Lane 1:

HEK293 whole cell lysate at 20 µg

Lane 2:

HEK293T whole cell lysate at 20 µg

Lane 3:

BirA-SLAP75 transfected HEK293T whole cell lysate, mock induced at 20 µg

Lane 4:

BirA-SLAP75 transfected HEK293T whole cell lysate, DOX induced at 20 µg

Predicted band size: 35 kDa

Observed band size: 75 kDa

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