Anti-PDIA6 antibody

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PDIA6 antibody (AB11432)

Immunocytochemistry/ Immunofluorescence analysis of 70% confluent log phase A549 cells labeling PDIA6 with ab11432. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ab11432 at 1/250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Secondary Antibody, Alexa Fluor^® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) were stained with SlowFade^® Gold Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Rhodamine Phalloidin, 1/300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PDIA6 antibody (AB11432)

ICC/IF image of ab11432 stained HeLa 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 (ab11432, 1µg/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.

• WB

Supplier Data

Western blot - Anti-PDIA6 antibody (AB11432)

Detected by chemiluminescence.

All lanes:

Western blot - Anti-PDIA6 antibody (ab11432) at 1/250 dilution

Lane 1:

A549 cell lysate at 30 µg

Lane 2:

A431 cell lysate at 30 µg

Lane 3:

HeLa cell lysate at 30 µg

Lane 4:

MCF7 cell lysate at 30 µg

Lane 5:

K-562 cell lysate at 30 µg

Lane 6:

Caco-2 cell lysate at 30 µg

Lane 7:

HepG2 cell lysate at 30 µg

Lane 8:

COLO 205 cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L), HRP conjugate at 1/2500 dilution

Predicted band size: 48 kDa

Observed band size: 48 kDa

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