Overlay histogram showing HAP1 wildtype (green line) and HAP1-CAT knockout cells (red line) stained with ab110292. The cells were fixed with 4% formaldehyde (10 min)  and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab110292, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) presorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.

A mouse IgG1 isotype control antibody  (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CAT  knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

Immunocytochemical staining of Human HeLa cells. The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were incubated with the ab110292 (5 µg/ml) for 2 hours at room temperature or over night at 4°C. The secondary antibody, Alexa Fluor® 488 goat anti-mouse IgG (green), was used at a 1/1000 dilution for 1 hour. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). The target protein locates mainly in peroxisomes.

Immunoprecipitation of Catalase using ab110292 from:
Lane 1: HepG2 cells
Lane 2: Cow heart sample
Lane 3: Rat liver sample

Predicted band size is 60 kDa.

Flow cytometric analysis of HL-60 cells using ab110292 at 1 µg/mL, (blue) or an isotype control (red).