Anti-XAF1 抗体

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-XAF1 antibody (AB81353)

ICC/IF image of ab81353 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab81353, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-XAF1 antibody (AB81353)

Overlay histogram showing SiHa cells stained with ab81353 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with ab81353 (1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight^®488 conjugated goat anti-rabbit IgG (5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

• WB

Supplier Data

Western blot - Anti-XAF1 antibody (AB81353)

Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours.

All lanes:

Western blot - Anti-XAF1 antibody (ab81353) at 0.5 µg/mL

All lanes:

Human Jurkat whole cell lysates at 30 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP secondary antibody at 1/5000 dilution

Predicted band size: 35 kDa

Observed band size: 35 kDa

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• WB

Supplier Data

Western blot - Anti-XAF1 antibody (AB81353)

All lanes:

Western blot - Anti-XAF1 antibody (ab81353) at 0.5 µg/mL

All lanes:

HEPG2 whole cell lysate at 40 µg

Predicted band size: 35 kDa

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