Recombinant Anti-Chd1 antibody [EPR23534-403] (ab303486)

Blocking and dilution buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS

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.

Blocking and dilution 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).

Blocking and dilution buffer and concentration: 5% NFDM/TBST

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