Anti-DR5 antibody

• ICC

Supplier Data

Immunocytochemistry - Anti-DR5 antibody (AB16329)

Immunocytochemistry analysis of DR5 was done on 70% confluent log phase HeLa cell. 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 ab16329 at 2 μg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ 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 Alexa Fluor^® 555 Rhodamine Phalloidin at 1/300. Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

• ICC

Supplier Data

Immunocytochemistry - Anti-DR5 antibody (AB16329)

Immunocytochemistry analysis of DR5 (TRAIL-R2) was performed using HCT 116 cells and HCT 116 treated with Thapsigargin (1 μM, 36 hours). The cells were fixed with 4% paraformaldehyde for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with ab16329 at 2 μg/mL in 0.1% BSA and incubated overnight at 4 degree and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 at a dilution of 1/2000 for 45 minutes at room temperature (Panel a : green) in HCT 116 treated cells. Nuclei (Panel b : blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI. F-actin (Panel c : red) was stained with Rhodamine Phalloidin 1/300. Panel d represents the merged image of HCT116 treated cells, which shows higher expression for TRAIL-R2 protein showing localization in nucleus, cytoplasm and membrane. Panel e represents the merged image of untreated HCT 116 cells, that shows lower or no expression for TRAIL-R2 protein. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

• WB

Supplier Data

Western blot - Anti-DR5 antibody (AB16329)

Detected by chemiluminescence

All lanes:

Western blot - Anti-DR5 antibody (ab16329) at 2 µg/mL

Lane 1:

A549 cell lysate at 20 µg

Lane 2:

HEL 92.1.7 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Superclonal™ at 1/2500 dilution

Predicted band size: 47 kDa

false

• WB

Supplier Data

Western blot - Anti-DR5 antibody (AB16329)

All lanes:

Western blot - Anti-DR5 antibody (ab16329) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 25 µg

Lane 2:

PC12 cell lysate at 25 µg

Secondary

All lanes:

HRP conjugated anti-rabbit

Predicted band size: 195 kDa,47 kDa

Observed band size: 58 kDa

true