Anti-Proteasome 20S beta 6 antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S beta 6 antibody (AB3331)

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 4°C 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S beta 6 antibody (AB3331)

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.

• WB

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Western blot - Anti-Proteasome 20S beta 6 antibody (AB3331)

Samples were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel and the resolved proteins were then transferred onto a Nitrocellulose membrane by iBlot® 2 Dry Blotting System. The blot was probed with the primary antibody (ab3331) and detected by chemiluminescence with secondary antibody using the iBright FL 1000. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit.

All lanes:

Western blot - Anti-Proteasome 20S beta 6 antibody (ab3331) at 1 µg/mL

Lane 1:

HeLa nuclear enriched extract lysate at 30 µg

Lane 2:

3T3-L1 nuclear enriched extract lysate at 30 µg

Lane 3:

PC-3 nuclear enriched extract lysate at 30 µg

Lane 4:

MCF7 nuclear enriched extract lysate at 30 µg

Lane 5:

A549 nuclear enriched extract lysate at 30 µg

Lane 6:

PANC-1 nuclear enriched extract lysate at 30 µg

Lane 7:

Mouse Kidney nuclear enriched extract lysate at 30 µg

Lane 8:

Mouse Liver nuclear enriched extract lysate at 30 µg

Lane 9:

Rat Liver nuclear enriched extract lysate at 30 µg

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/6000 dilution

Predicted band size: 25 kDa

Observed band size: 21 kDa

true

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S beta 6 antibody (AB3331)

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 4°C 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S beta 6 antibody (AB3331)

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 4°C 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.

• WB

Supplier Data

Western blot - Anti-Proteasome 20S beta 6 antibody (AB3331)

All lanes:

Western blot - Anti-Proteasome 20S beta 6 antibody (ab3331) at 1/1000 dilution

Lane 1:

HeLa (Human epithelial adenocarcinoma cell line) whole cell lysate at 25 µg

Lane 2:

BAEC (Bovine aortic endothelial cell line) whole cell lysate at 25 µg

Secondary

All lanes:

HRP-conjugated secondary antibody

Predicted band size: 25 kDa

false

• WB

Supplier Data

Western blot - Anti-Proteasome 20S beta 6 antibody (AB3331)

All lanes:

Western blot - Anti-Proteasome 20S beta 6 antibody (ab3331) at 1 µg/mL

Lane 1:

Nuclear enriched extracts from untransfected PC-3 cells

Lane 2:

Nuclear enriched extracts from non-targeting scrambled siRNA transfected PC-3 cells

Lane 3:

Nuclear enriched extracts from PSMB6 knockdown PC-3 cells

Secondary

All lanes:

Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP at 1/6000 dilution

Predicted band size: 25 kDa

false