All lanes : Anti-Catalase antibody [EPR20198] (ab209211) at 1/2000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CAT knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Lanes 1-2: Merged signal (red and green). Green - ab209211 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab209211 Anti-Catalase antibody [EPR20198] was shown to specifically react with Catalase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265250 (knockout cell lysate ab256859) was used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab209211 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 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-Catalase antibody [EP1929Y] - Peroxisome Marker (ab76024) at 1/10000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CAT knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 60 kDa
Observed band size: 60 kDa

Lanes 1-2: Merged signal (red and green). Green - ab76024 observed at 60 kDa. Red - loading control ab8245 observed at 37 kDa.

 ab76024 Anti-Catalase antibody [EP1929Y] - Peroxisome Marker was shown to specifically react with Catalase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265250 (knockout cell lysate ab256859) was used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab76024 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: 4 bp deletion in exon 1.

 

Allele-2: 1 bp deletion in exon 1.

 

Allele-3: Insertion of the selection cassette in exon 1.

 

Representative images of CAT knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa 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.