Anti-Gli1 antibody [EPR4523]

• WB

Lab

Western blot - Anti-Gli1 antibody [EPR4523] (AB134906)

Western blot : Rabbit Monoclonal [EPR4523] to Gli1 ab134906 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 145 kDa in Wild-type A549 cell lysates with no signal observed at this size in GLI1 knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Gli1 antibody [EPR4523] (ab134906) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

Human GLI1 knockout A549 cell line at 20 µg

Lane 2:

Western blot - Human GLI1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-gli1-knockout-a549-cell-line-ab277815'>ab277815</a>) at 20 µg

Lane 3:

SH-SY5Y cell lysate at 40 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 118 kDa

Observed band size: 145 kDa

false

• WB

Supplier Data

Western blot - Anti-Gli1 antibody [EPR4523] (AB134906)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

The expression level of Gli1 in HepG2 and RD are consistent with describes in PMID : 19189974 and PMID : 32493277.

Negative control : HepG2 and RD (PMID : 19189974 and PMID : 32493277).

All lanes:

Western blot - Anti-Gli1 antibody [EPR4523] (ab134906) at 1/1000 dilution

Lane 1:

A673 (human muscle Ewing's Sarcoma) whole cell lysate at 20 µg

Lane 2:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

RD (Human muscle rhabdomyosarcoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 118 kDa

Observed band size: 160 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Gli1 antibody [EPR4523] (AB134906)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Gli1 antibody [EPR4523] (ab134906) at 1/1000 dilution

Lane 1:

RMS 13 (human rhabdomyosarcoma fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

RMS 13 (human rhabdomyosarcoma fibroblast) transfected with siRNA specifically targeting Gli1 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 118 kDa

Observed band size: 150 kDa

true

Exposure time: 180s