Anti-SEMA3B antibody

• WB

Lab

Western blot - Anti-SEMA3B antibody (AB48197)

Western blot : Anti-SEMA3B antibody (ab48197) staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab48197 was shown to bind specifically to SEMA3B. A band was observed at 81 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in SEMA3B knockout cell line (ab326024). Arrow indicates the target band.To generate this image, wild-type and SEMA3B knockout HCT 116 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-SEMA3B antibody (ab48197) at 1/5000 dilution

Lane 1:

Wild-type HCT 116 cell lysate at 20 µg

Lane 2:

Western blot - Human SEMA3B knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-sema3b-knockout-hct116-cell-line-ab326024'>ab326024</a>) at 20 µg

Lane 3:

A549 cell lysate at 20 µg

Lane 4:

RKO 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: 81 kDa

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