All lanes : Anti-Proteasome 20S beta 6 antibody (ab3331) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial adenocarcinoma cell line) whole cell lysate
Lane 2 : BAEC (Bovine aortic endothelial cell line) whole cell lysate

Lysates/proteins at 25 µg per lane.

Secondary
All lanes : HRP-conjugated secondary antibody

Predicted band size: 25 kDa

Immunocytochemistry/Immunofluorescence analysis of Proteasome 20S beta 6 (green) showing staining in the cytoplasm and nucleus of A431 (Human epidermoid carcinoma cell line) cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab3331 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunofluorescent analysis of Proteasome 20S Y (green) in NIH/3T3 (Mouse embryo fibroblast cell line) cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes and blocked with 3% Blocker BSA in PBS for 30 minutes at room temperature. Cells were stained with or without Proteasome 20S Y rabbit polyclonal antibody, at a concentration of 5 µg/mL for 1 hour at room temperature, and then incubated with a Goat anti-Rabbit (H+L) Superclonal Secondary Antibody, Alexa Fluor® 488 conjugate at 1/1000 dilution for 1 hour at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

Immunocytochemistry/Immunofluorescence analysis of Proteasome 20S beta 6 (green) showing staining in the cytoplasm and nucleus of BAEC (Bovine aortic endothelial cell line) cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab3331 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Immunocytochemistry/Immunofluorescence analysis of Proteasome 20S beta 6 (green) showing staining in the cytoplasm and nucleus of HeLa (Human epithelial adenocarcinoma cell line) cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ab3331 in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.