Anti-ILKAP antibody [EPR16145]

• IP

Supplier Data

Immunoprecipitation - Anti-ILKAP antibody [EPR16145] (AB196013)

Immunoprecipitation analysis of 293 (Human epithelial cells from embryonic kidney) whole cell extract labeling ILKAP using ab196013 at 1/30 dilution (Lane 2).
Lane 3 : IP using Rabbit monoclonal IgG (ab172730) instead of ab196013 in 293 (Human epithelial cells from embryonic kidney) whole cell extract.
Lane 1 : Input : 10 μg 293 (Human epithelial cells from embryonic kidney) whole cell extract.
Subsequent WB detection was performed using ab196013 at 1/1000 dilution.
An Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 was used as secondary antibody.

All lanes:

Immunoprecipitation - Anti-ILKAP antibody [EPR16145] (ab196013)

Predicted band size: 43 kDa

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• WB

Lab

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

Lanes 1-4 : Merged signal (red and green). Green - ab196013 observed at 49 kDa. Red - loading control ab8245 observed at 36 kDa.

ab196013 Anti-ILKAP antibody [EPR16145] was shown to specifically react with ILKAP in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266475 (knockout cell lysate ab258000) was used. Wild-type and ILKAP knockout samples were subjected to SDS-PAGE. ab196013 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ILKAP knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ILKAP knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ilkap-knockout-hek-293t-cell-line-ab266475'>ab266475</a>)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

HAP1 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 43 kDa

Observed band size: 49 kDa

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• WB

Lab

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : ILKAP knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lane 4 : HEK293 cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab196013 observed at 43 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab196013 was shown to recognize ILKAP when ILKAP knockout samples were used, along with additional cross-reactive bands. Wild-type and ILKAP knockout samples were subjected to SDS-PAGE. ab196013 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013) at 1/1000 dilution

All lanes:

Rat brain lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013) at 1/5000 dilution

All lanes:

Mouse brain lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013) at 1/20000 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa

false

• WB

Supplier Data

Western blot - Anti-ILKAP antibody [EPR16145] (AB196013)

All lanes:

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013) at 1/20000 dilution

All lanes:

293 (Human epithelial cells from embryonic kidney) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa

false