Anti-CHD8 antibody

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CHD8 antibody (AB224830)

Formalin-fixed, paraffin-embedded human ovarian carcinoma tissue stained for CHD8 using ab224830 at 1/200 dilution in immunohistochemical analysis. Detection : DAB staining.

The two panels show staining with different lots.

• IP

Supplier Data

Immunoprecipitation - Anti-CHD8 antibody (AB224830)

CHD8 was immunoprecipitated from HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate prepared using NETN buffer with ab224830 at 6 μg per reaction. Western blot was performed from the immunoprecipitate using ab224830 at 0.1 μg/ml.

Lane 1 : ab224830 IP in HEK-293T whole cell lysate.

Lane 2 : ab224830 IP in HEK-293T whole cell lysate.

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

Detection : Chemiluminescence with exposure time of 3 minutes.

All lanes:

Immunoprecipitation - Anti-CHD8 antibody (ab224830)

Predicted band size: 291 kDa

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• WB

Supplier Data

Western blot - Anti-CHD8 antibody (AB224830)

Lysates prepared using NETN buffer.

All lanes:

Western blot - Anti-CHD8 antibody (ab224830) at 0.1 µg/mL

Lane 1:

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

Lane 2:

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

Lane 3:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 50 µg

Lane 4:

TCMK-1 (mouse kidney epithelial cell line) whole cell lysate at 50 µg

Lane 5:

NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 50 µg

Predicted band size: 291 kDa

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Exposure time: 3min

• WB

Lab

Western blot - Anti-CHD8 antibody (AB224830)

Western blot : Anti-CHD8 antibody (ab224830) staining at 1/5000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab224830 was shown to bind specifically to CHD8. A band was observed at 355 kDa in wild-type A549 cell lysates with no signal observed at this size in CHD8 knockout cell line. To generate this image, wild-type and CHD8 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-CHD8 antibody (ab224830) at 1/5000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

CHD8 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HEK-293T 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: 355 kDa

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