Anti-Catalase antibody - Peroxisome Marker (ab52477) at 1/1000 dilution + TF1 lysate at 10 µg

Secondary
Goat anti-rabbit HRP labeled at 1/2000 dilution

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

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Catalase knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab52477 observed at 60 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab52477 was shown to specifically react with Catalase when Catalase knockout samples were used. Wild-type and Catalase knockout samples were subjected to SDS-PAGE. ab52477 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

IHC image of Catalasestaining in a formalin fixed, paraffin embedded normal human liver tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab52477, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

 

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

All lanes : Anti-Catalase antibody - Peroxisome Marker (ab52477) at 1 µg/ml

Lane 1 : Whole cell lysate prepared from 293T cells
Lane 2 : Whole cell lysate prepared from HeLa cells
Lane 3 : Whole cell lysate prepared from U251 cells
Lane 4 : Whole cell lysate prepared from Jurkat cells
Lane 5 : Whole cell lysate prepared from U2OS cells

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution

Developed using the ECL technique.

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


Exposure time: 5 minutes

See Abreview

All lanes : Anti-Catalase antibody - Peroxisome Marker (ab52477) at 1 µg/ml

Lane 1 : Whole cell lysate prepared from 9L cells
Lane 2 : Whole cell lysate prepared from PC12 cells
Lane 3 : Whole cell lysate prepared from RIE-1 cells

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP conjugated pig anti-rabbit polyclonal at 1/5000 dilution

Developed using the ECL technique.

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


Exposure time: 5 minutes

See Abreview