Anti-Peroxiredoxin 1/PAG antibody (ab41906)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Peroxiredoxin 1/PAG knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: Jurkat cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab41906 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab41906 was shown to recognize Peroxiredoxin 1/PAG when Peroxiredoxin 1/PAG knockout samples were used, along with additional cross-reactive bands. Wild-type and Peroxiredoxin 1/PAG knockout samples were subjected to SDS-PAGE. ab41906 and ab8245 (loading control to GAPDH) at a concentration of 1 µg/ml  and diluted to  1/10000 respectively were 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 hour at room temperature before imaging.

IHC image of Peroxiredoxin 1/PAG staining in human liver carcinoma FFPE section, performed on a Bond^TM 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 ab41906, 1µg/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.

ab41906 was shown to react with PRDX1 in wild-type U-2 OS cells in Western blot with loss of signal observed in a PRDX1 knockout cell line. Wild-type U-2 OS and PRDX1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab41906 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2ug/mL before imaging. This data was kindly provided by the YCharOS Inc., an open science company with the mission of characterizing every commercially available antibody reagent. Abcam are working with YCharOS to support their mission of antibody characterisation using knockout cell lines.