The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
1/2000 - 1/4000. Detects a band of approximately 26 kDa (predicted molecular weight: 22 kDa).
Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
Use a concentration of 200 mg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Involved in redox regulation of the cell. Reduces peroxides with reducing equivalents provided through the thioredoxin system. It is not able to receive electrons from glutaredoxin. May play an important role in eliminating peroxides generated during metabolism. Might participate in the signaling cascades of growth factors and tumor necrosis factor-alpha by regulating the intracellular concentrations of H(2)O(2).
Belongs to the ahpC/TSA family. Contains 1 thioredoxin domain.
Western blot - Anti-Peroxiredoxin 2 antibody [1E8] (ab50862)
Predicted band size : 22 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: Peroxiredoxin 2 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 - ab50862 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa. ab50862 was shown to specifically react with Peroxiredoxin 2 when Peroxiredoxin 2 knockout samples were used. Wild-type and Peroxiredoxin 2 knockout samples were subjected to SDS-PAGE. ab50862 and ab181602 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG (H + L) (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777)secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
Western blot - Peroxiredoxin 2 antibody [1E8] (ab50862)
All lanes : Anti-Peroxiredoxin 2 antibody [1E8] (ab50862) at 1/2000 dilution
Lane 1 : HeLa cell lysate Lane 2 : 293T cell lysate Lane 3 : SH-SY5Y cell lysate Lane 4 : HepG2 cell lysate
Overlay histogram showing HeLa cells stained with ab50862 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab56802, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.