Anti-Chd1 antibody [EPR23534-403] - BSA and Azide free

• WB

Lab

Western blot - Anti-Chd1 antibody [EPR23534-403] - BSA and Azide free (AB303487)

This data was developed using ab303486, the same antibody clone in a different buffer formulation.

The identity of the bands between 40 kDa and 100 kDa are non-specific.

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

False colour image of Western blot : Anti-Chd1 antibody [EPR23534-403] (ab303486) 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, ab303486 was shown to bind specifically to Chd1. A band was observed at 250 kDa in wild-type HEK-293 cell lysates whereas no signal observed at this size in HEK-293 knockout cell line ab266512 (knockout cell lysate 263151). To generate this image, wild-type and Chd1 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-Chd1 antibody [EPR23534-403] (<a href='/en-us/products/primary-antibodies/chd1-antibody-epr23534-403-ab303486'>ab303486</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293 (human embryonic kidney epithelial cell), whole cell lysate

Lane 2:

Chd1 knockout HEK-293 whole cell lysate

Lane 3:

LNCaP (human prostate carcinoma epithelial cell), whole cell lysate

Lane 4:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/100000 dilution

Observed band size: 250 kDa

false

• WB

Lab

Western blot - Anti-Chd1 antibody [EPR23534-403] - BSA and Azide free (AB303487)

This data was developed using ab303486, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab303486 was shown to bind specifically to Chd1. A band was observed at 250 kDa in wide-type HEK-293T cell lysates with no signal observed at this size in Chd1 knockout cell line ab266512 (knockout cell lysate 263151).

All lanes:

Western blot - Anti-Chd1 antibody [EPR23534-403] (<a href='/en-us/products/primary-antibodies/chd1-antibody-epr23534-403-ab303486'>ab303486</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell), cell lysate at 20 µg

Lane 2:

Chd1 knockout HEK-293T, cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 250 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Chd1 antibody [EPR23534-403] - BSA and Azide free (AB303487)

This data was developed using ab303486, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lysates were freshly made and used immediately to minimize protein degradation.

All lanes:

Western blot - Anti-Chd1 antibody [EPR23534-403] (<a href='/en-us/products/primary-antibodies/chd1-antibody-epr23534-403-ab303486'>ab303486</a>) at 1/1000 dilution

All lanes:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 250 kDa

false

Exposure time: 15s