Anti-Proteasome 20S LMP7 antibody

• ICC

Supplier Data

Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (AB3329)

Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Proteasome 20S LMP7 with ab3329 at 5μg/ml. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with or without Anti-Proteasome 20S LMP7 antibody (ab3329), at a concentration of 5μg/ml for 1 hour at room temperature, and then incubated with a Alexa Fluor^® 488 goat anti-rabbit IgG secondary antibody at a dilution of 1/1000 for 1 hour s at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.

• ICC

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Immunocytochemistry - Anti-Proteasome 20S LMP7 antibody (AB3329)

ICC/IF image of ab3329 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 (ab3329, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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

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Western blot - Anti-Proteasome 20S LMP7 antibody (AB3329)

Western blot of proteasome 20S LMP7 from HeLa cell extract using ab3329.

All lanes:

Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329)

Predicted band size: 30 kDa

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• WB

Supplier Data

Western blot - Anti-Proteasome 20S LMP7 antibody (AB3329)

Detected by chemiluminescence.

All lanes:

Western blot - Anti-Proteasome 20S LMP7 antibody (ab3329) at 2 µg/mL

Lane 1:

HeLa whole cell extracts at 30 µg

Lane 2:

HeLa treated with IFN gamma (100ng/ml IFN gamma for 72h) whole cell extracts at 30 µg

Lane 3:

U-937 whole cell extracts at 30 µg

Lane 4:

Raji whole cell extracts at 30 µg

Lane 5:

Ramos whole cell extracts at 30 µg

Secondary

All lanes:

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

Predicted band size: 30 kDa

Observed band size: 20 kDa

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