All lanes : Anti-PCBD1 antibody [EPR9760(B)] (ab138518) at 1/500 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : PCBD1 knockout HEK293T cell lysate
Lane 3 : Caco-2 cell lysate
Lane 4 : Human ovary cancer tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 12 kDa
Observed band size: 12 kDa

Lanes 1-4: Merged signal (red and green). Green - ab138518 observed at 12 kDa. Red - loading control ab8245 observed at 36 kDa.

 ab138518 Anti-PCBD1 antibody [EPR9760(B)]  was shown to specifically react with PCBD1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266261 (knockout cell lysate ab258095) was used. Wild-type and PCBD1 knockout samples were subjected to SDS-PAGE. ab138518 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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.

Homozygous: Insertion of the selection cassette in exon2

Representative images of PCBD1 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 an EVOS M5000 microscope.