Anti-BioID2 antibody [SS 3A5-E2] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BioID2 antibody [SS 3A5-E2] - BSA and Azide free (AB269568)

ab232733 staining BioID2 in HeLa cells expressing DOX-inducible BioID2 dystonia mutant of TorsinA. Parental HeLa cells have been used as negative control. The cells were fixed with 100% methanol (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 ab232733 at 1 μg/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 ab150084, 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab232733).

• WB

Lab

Western blot - Anti-BioID2 antibody [SS 3A5-E2] - BSA and Azide free (AB269568)

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 3% milk before ab232733 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab232733).

All lanes:

Western blot - Anti-BioID2 antibody [SS 3A5-E2] (<a href='/en-us/products/primary-antibodies/bioid2-antibody-ss-3a5-e2-ab232733'>ab232733</a>) at 1 µg/mL

Lane 1:

HeLa whole cell lysate at 10 µg

Lane 2:

BiolD2-TorsinA transfected HeLa whole cell lysate, non-induced at 10 µg

Lane 3:

BiolD2-TorsinA transfected HeLa whole cell lysate, DOX induced at 10 µg

Observed band size: 70 kDa

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