Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Peroxiredoxin 2/PRP knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: LnCaP cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133481 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133481 was shown to specifically react with Peroxiredoxin 2/PRP when Peroxiredoxin 2/PRP knockout samples were used. Wild-type and Peroxiredoxin 2/PRP knockout samples were subjected to SDS-PAGE. ab133481 and ab8245 (loading control to GAPDH) were diluted 1/50 000 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)
and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

Overlay histogram showing HeLa cells stained with ab133481 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133481, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Immunofluorescence analysis of HeLa cells labelling Peroxiredoxin 2/PRP with ab133481 at 1/100 dilution.

All lanes : Anti-Peroxiredoxin 2/PRP antibody [EPR5155] (ab133481) at 1/5000 dilution

Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : PRDX2 knockout HEK-293T cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 22 kDa
Observed band size: 22 kDa

Lanes 1- 2: Merged signal (red and green). Green - ab133481 observed at 22 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

 ab133481 was shown to react with Peroxiredoxin 2/PRP in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266392 (knockout cell lysate ab257041) was used. Wild-type HEK-293T and PRDX2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133481 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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-Peroxiredoxin 2/PRP antibody [EPR5155] (ab133481) at 1/50000 dilution

Lane 1 : 293T cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : LnCaP cell lysate
Lane 4 : U937 cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 22 kDa