Anti-GABARAP antibody - N-terminal

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-GABARAP antibody - N-terminal (AB227732)

Ice-cold methanol-fixed U-2 OS (human bone osteosarcoma epithelial cell line) cells stained for GABARAP (green) using ab227732 at 1/500 dilution in ICC/IF.

Nuclear counterstain : Hoechst 33342 (blue).

• WB

Supplier Data

Western blot - Anti-GABARAP antibody - N-terminal (AB227732)

15% SDS-PAGE gel.

All lanes:

Western blot - Anti-GABARAP antibody - N-terminal (ab227732) at 1/2000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

Lane 2:

A431 (human epidermoid carcinoma cell line) whole cell lysate at 30 µg

Lane 3:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 30 µg

Predicted band size: 13 kDa

false

• WB

Supplier Data

Western blot - Anti-GABARAP antibody - N-terminal (AB227732)

15% SDS-PAGE gel.

All lanes:

Western blot - Anti-GABARAP antibody - N-terminal (ab227732) at 1/1000 dilution

Lane 1:

Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell extract at 30 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GABARAP shRNA, whole cell extract at 30 µg

Predicted band size: 13 kDa

false

• WB

Supplier Data

Western blot - Anti-GABARAP antibody - N-terminal (AB227732)

15% SDS-PAGE gel.

All lanes:

Western blot - Anti-GABARAP antibody - N-terminal (ab227732) at 1/2000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 30 µg

Lane 2:

Raji (human Burkitt's lymphoma cell line) whole cell lysate at 30 µg

Lane 3:

NCI-H929 whole cell lysate at 30 µg

Predicted band size: 13 kDa

false

• WB

Lab

Western blot - Anti-GABARAP antibody - N-terminal (AB227732)

Western blot : Anti-GABARAP antibody (ab227732) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab227732 was shown to bind specifically to GABARAP. A band was observed at 15 kDa in wild-type A549 cell lysates with no signal observed at this size in GABARAP knockout cell line. To generate this image, wild-type and GABARAP knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-GABARAP antibody - N-terminal (ab227732) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

GABARAP knockout A549 cell lysate at 20 µg

Lane 3:

T-47D cell lysate at 20 µg

Lane 4:

A549 Nuclear cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 15 kDa

false

• WB

Supplier Data

Western blot - Anti-GABARAP antibody - N-terminal (AB227732)

15% SDS-PAGE gel.

All lanes:

Western blot - Anti-GABARAP antibody - N-terminal (ab227732) at 1/10000 dilution

Lane 1:

Non-transfected HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 30 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with GABARAP whole cell lysate at 30 µg

Predicted band size: 13 kDa

false