Anti-Cytokeratin 7 antibody [EP1620Y] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-Cytokeratin 7 antibody [EP1620Y] - BSA and Azide free (AB181832)

This data was developed using ab68460, the same antibody clone in a different buffer formulation.

ab68460 (purified) at 1 : 20 dilution (0.6μg) immunoprecipitating Cytokeratin 7 in T-47D whole cell lysate.
Lane 1 (input) : T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab68460 & T-47D whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab68460 in T-47D whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-Cytokeratin 7 antibody [EP1620Y] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-ep1620y-ab68460'>ab68460</a>)

Predicted band size: 51 kDa

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• WB

Unknown

Western blot - Anti-Cytokeratin 7 antibody [EP1620Y] - BSA and Azide free (AB181832)

This data was developed using ab68460, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

~45 kDa bands are supported by the literature and competitor's products, which may be an isoform (PMID : 16265353).

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EP1620Y] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-ep1620y-ab68460'>ab68460</a>) at 1/20000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg/mL

Lane 2:

T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 20 µg/mL

Lane 3:

SK-BR-3 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg/mL

Lane 4:

Human placenta tissue lysate at 20 mg/mL

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 51 kDa

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• WB

Unknown

Western blot - Anti-Cytokeratin 7 antibody [EP1620Y] - BSA and Azide free (AB181832)

This data was developed using ab68460, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EP1620Y] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-ep1620y-ab68460'>ab68460</a>) at 1/20000 dilution

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

HRP conjugated Goat anti-Rabbit at 1/2000 dilution

Predicted band size: 51 kDa

Observed band size: 51 kDa

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• WB

Lab

Western blot - Anti-Cytokeratin 7 antibody [EP1620Y] - BSA and Azide free (AB181832)

This data was developed using ab68460, the same antibody clone in a different buffer formulation. Western blot : Anti-KRT7 antibody [EP1620Y] (ab68460) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab68460 was shown to bind specifically to KRT7. A band was observed at 40-55 kDa in wild-type A549 cell lysates with no signal observed at this size in KRT7 knockout cell line. To generate this image, wild-type and KRT7 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-Cytokeratin 7 antibody [EP1620Y] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-ep1620y-ab68460'>ab68460</a>) at 1/10000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

KRT7 knockout A549 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MCF7 cell lysate at 20 µg

Observed band size: 40-55 kDa

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