Anti-Robo1 antibody

• WB

Lab

Western blot - Anti-Robo1 antibody (AB245516)

Western blot : Anti-ROBO1 antibody (ab245516) staining at 1/2000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab245516 was shown to bind specifically to ROBO1. A band was observed at 200 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in ROBO1 knockout cell line. To generate this image, wild-type and ROBO1 knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-Robo1 antibody (ab245516) at 1/2000 dilution

Lane 1:

Wild-type U-87 MG cell lysate at 20 µg

Lane 2:

ROBO1 knockout U-87 MG cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 200 kDa

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• IP

Supplier Data

Immunoprecipitation - Anti-Robo1 antibody (AB245516)

Robo1 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate using ab245516 at 3 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab245516 at 1 μg/ml.

Lane 1 : ab245516 IP in HeLa whole cell lysate.
Lane 2 : Control IgG in HeLa whole cell lysate.

Detection : Chemiluminescence with an exposure time of 3 seconds.

All lanes:

Immunoprecipitation - Anti-Robo1 antibody (ab245516)

Predicted band size: 181 kDa

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• WB

Supplier Data

Western blot - Anti-Robo1 antibody (AB245516)

All lanes:

Western blot - Anti-Robo1 antibody (ab245516) at 0.04 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Lane 4:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

Predicted band size: 181 kDa

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Exposure time: 10s