Anti-Chk2 antibody [EPR19482] - BSA and Azide free

• WB

Lab

Western blot - Anti-Chk2 antibody [EPR19482] - BSA and Azide free (AB251477)

This data was developed using the same antibody clone in a different buffer formulation (ab207446). False colour image of Western blot : Anti-Chk2 antibody [EPR19482] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab207446 was shown to bind specifically to Chk2. A band was observed at 67 kDa in wild-type A549 cell lysates with no signal observed at this size in CHEK2 knockout cell line ab276098 (knockout cell lysate ab276098). To generate this image, wild-type and CHEK2 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.

Lanes 1 - 4:

Western blot - Anti-Chk2 antibody [EPR19482] (<a href='/en-us/products/primary-antibodies/chk2-antibody-epr19482-ab207446'>ab207446</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Anti-Chk2 antibody [EPR19482] - BSA and Azide free (ab251477) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human CHEK2 (Chk2) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-chek2-chk2-knockout-a549-cell-line-ab276098'>ab276098</a>)

Lane 2:

CHEK2 knockout A549 cell lysate at 20 µg

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

MDA-MB-231 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: 67 kDa

false

• WB

Lab

Western blot - Anti-Chk2 antibody [EPR19482] - BSA and Azide free (AB251477)

This data was developed using the same antibody clone in a different buffer formulation (ab207446).

Lanes 1-4 : Merged signal (red and green). Green - ab207446 observed at 68 kDa. Red - loading control ab8245 observed at 37 kDa.

ab207446 Anti-Chk2 antibody [EPR19482] was shown to specifically react with Chk2 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264815 (knockout cell lysate ab257104) was used. Wild-type and Chk2 knockout samples were subjected to SDS-PAGE. ab207446 and Anti-alpha Tubulin antibody [EP1332Y] - Microtubule Marker (ab52866) 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-Chk2 antibody [EPR19482] (<a href='/en-us/products/primary-antibodies/chk2-antibody-epr19482-ab207446'>ab207446</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CHEK2 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CHEK2 (Chk2) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-chek2-chk2-knockout-hela-cell-line-ab264815'>ab264815</a>)

Lane 3:

HEK-293 cell lysate at 20 µg

Lane 4:

MDA-MB-231 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 1/20000 dilution

Predicted band size: 61 kDa

Observed band size: 68 kDa

false