Human ALDH1A1 knockout A549 cell lysate

• WB

Lab

Western blot - Human ALDH1A1 knockout A549 cell lysate (AB261673)

Western blot : Anti-ALDH1A1 antibody [ALDH1A1/1381] (ab219303) staining at 1 ug/ml, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab219303 was shown to bind specifically to ALDH1A1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in ALDH1A1 knockout cell line. To generate this image, wild-type and ALDH1A1 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-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-ALDH1A1 antibody [ALDH1A1/1381] (<a href='/en-us/products/primary-antibodies/aldh1a1-antibody-aldh1a1-1381-ab219303'>ab219303</a>) at 1 µg/mL

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ALDH1A1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-aldh1a1-knockout-a549-cell-line-ab261864'>ab261864</a>)

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell lysate (ab261673)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

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• WB

Lab

Western blot - Human ALDH1A1 knockout A549 cell lysate (AB261673)

Western blot : Anti-ALDH1A1 antibody [EP1933Y] (ab52492) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52492 was shown to bind specifically to ALDH1A1. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in ALDH1A1 knockout cell line. To generate this image, wild-type and ALDH1A1 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-ALDH1A1 antibody [EP1933Y] - C-terminal (<a href='/en-us/products/primary-antibodies/aldh1a1-antibody-ep1933y-c-terminal-ab52492'>ab52492</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

ALDH1A1 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-aldh1a1-knockout-a549-cell-line-ab261864'>ab261864</a>)

Lane 2:

Western blot - Human ALDH1A1 knockout A549 cell lysate (ab261673)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

MCF7 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

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• NGS

Supplier Data

Next Generation Sequencing - Human ALDH1A1 knockout A549 cell lysate (AB261673)

Knockout achieved by CRISPR/Cas9; X = 8 bp deletion; Frameshift : 98%