All lanes : Anti-HDAC2 antibody [EPR20117] (ab219053) at 1/1000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab219053 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab219053 Anti-HDAC2 antibody [EPR20117] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab219053 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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 : Anti-HDAC2 antibody [EPR5001] - ChIP Grade (ab124974) at 1/10000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HDAC2 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 55 kDa
Observed band size: 60 kDa why is the actual band size different from the predicted?

Lanes 1-2: Merged signal (red and green). Green - ab124974 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab124974 Anti-HDAC2 antibody [EPR5001] was shown to specifically react with HDAC2 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266590 (knockout cell lysate ab256939) was used. Wild-type and HDAC2 knockout samples were subjected to SDS-PAGE. ab124974 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 10000 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.

 

Allele-1: 11 bp deletion in exon2

 

Allele-2: 1 bp insertion in exon 2.

 

Representative images of HDAC2 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.