All lanes : Anti-Drosha antibody [EPR12794] (ab183732) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : DROSHA knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 159 kDa

Lanes 1-3: Merged signal (red and green). Green - ab183732 observed at 159 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab183732 Anti-Drosha antibody [EPR12794] was shown to specifically react with Drosha in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266217 (knockout cell lysate ab257171) was used. Wild-type and Drosha knockout samples were subjected to SDS-PAGE. ab183732 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 10000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 1 bp insertion in exon 4

Representative images of DROSHA 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.