The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000 - 1/50000. Detects a band of approximately 22 kDa (predicted molecular weight: 22 kDa).
1/250 - 1/500. Perform antigen retrieval
1/10 - 1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
1/100 - 1/250.
Application notesIs unsuitable for IP.
FunctionInvolved in redox regulation of the cell. Reduces peroxides with reducing equivalents provided through the thioredoxin system but not 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). Reduces an intramolecular disulfide bond in GDPD5 that gates the ability to GDPD5 to drive postmitotic motor neuron differentiation.
Sequence similaritiesBelongs to the ahpC/TSA family. Contains 1 thioredoxin domain.
Post-translational modificationsPhosphorylated on Thr-90 during the M-phase, which leads to a more than 80% decrease in enzymatic activity.
Cellular localizationCytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Western blot - Anti-Peroxiredoxin 1 antibody [EPR5433] (ab109498)
Predicted band size : 22 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: Peroxiredoxin 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 - ab109498 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109498 was shown to specifically react with Peroxiredoxin when Peroxiredoxin knockout samples were used. Wild-type and Peroxiredoxin knockout samples were subjected to SDS-PAGE. ab109498 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 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 1/10 000 dilution for 1 h at room temperature before imaging.
Overlay histogram showing HeLa cells stained with ab109498 (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 (ab109498, 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) (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.
Western blot - Peroxiredoxin 1 antibody [EPR5433] (ab109498)
All lanes : Anti-Peroxiredoxin 1 antibody [EPR5433] (ab109498) at 1/10000 dilution
Lane 1 : MCF-7 cell lysate Lane 2 : 293T cell lysate Lane 3 : K562 cell lysates
Lysates/proteins at 10 µg per lane.
Secondary Standard HRP labelled goat anti-rabbit at 1/2000 dilution
Predicted band size : 22 kDa Observed band size : 22 kDa