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.

 

 

Immunohistochemical analysis of Paraffin-embedded human bladder cancer tissue sections labeling Catalase with ab76024 at 1/1000. Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Sections were counterstained with Hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).

Granular cytoplasmic staining on human bladder cancer.

All lanes : Anti-Catalase antibody [EP1929Y] - Peroxisome Marker (ab76024)

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : CAT knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 60 kDa

Lanes 1 - 3: Merged signal (red and green). Green - ab76024 observed at 60 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab76024 was shown to specifically react with CAT when CAT knockout samples were used. Wild-type and CAT knockout samples were subjected to SDS-PAGE. Ab76024 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) labeling Catalase ab76024 at 1/100. Cells were fixed with 100% Methanol. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. 

ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/100 was used as counterstain antibody.

Confocal image showing membranous staining in HeLa cells.

 

Anti-Catalase antibody [EP1929Y] - Peroxisome Marker (ab76024) at 1/20000 dilution + HeLa cell lysate at 10 µg

Secondary
HRP labelled goat anti-rabbit at 1/2000 dilution

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

Immunohistochemical staining of Catalase in paraffin embedded human normal brain tissue using ab76024 at a 1/100 dilution.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.