Product nameAnti-Peroxiredoxin 1/PAG antibody
See all Peroxiredoxin 1/PAG primary antibodies
DescriptionRabbit polyclonal to Peroxiredoxin 1/PAG
SpecificityThis antibody detects Peroxiredoxin 1 protein in human samples. The antibody is specific for Peroxiredoxin 1/PAG and shows no cross reactivity with other Prx isoforms.
Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
Species reactivityReacts with: Mouse, Rat, Human, African green monkey
Predicted to work with: Cow, Chinese hamster
- Recombinant human Peroxiredoxin 1/PAG protein (ab74172) can be used as a positive control in WB. TSU Pr1 cells for cell staining, human PC3 cell lysate for western blotting.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.05% Sodium azide
Our Abpromise guarantee covers the use of ab15571 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use at an assay dependent concentration.|
|WB||1/1000. Detects a band of approximately 20 kDa (predicted molecular weight: 22 kDa).|
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.
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.
- Information by UniProt
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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 - ab15571 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab15571 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. ab15571 and ab8245 (loading control to GAPDH) were diluted to 1/1000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
ICC/IF image of ab15571 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab15571, 1/200 dilution) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
IHC image of ab15571 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab15571, 1/100 dilution, 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.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
All lanes : Anti-Peroxiredoxin 1/PAG antibody (ab15571) at 1/1000 dilution
Lane 1 : Cell line indicated at 25 µg
All lanes : HRP-conjugated Goat anti-rabbit at 1/20000 dilution
Predicted band size: 22 kDa
Observed band size: 20 kDa why is the actual band size different from the predicted?
Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.
Western blot analysis of Peroxiredoxin 1//PAG was performed by loading 25ug of various whole cell lysates onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% Milk/TBST for at least 1 hour. Membranes were probed with ab15571 overnight at 4°C on a rocking platform. Membranes were washed in TBS-0.1%Tween 20 and probed with a goat anti-rabbit-HRP secondary antibody for at least one hour. Membranes were washed and chemiluminescent detection performed.
This product has been referenced in:
- Yang W et al. Neutrophils promote the development of reparative macrophages mediated by ROS to orchestrate liver repair. Nat Commun 10:1076 (2019). Read more (PubMed: 30842418) »
- He Y et al. Circulating Peroxiredoxin-1 is a novel damage-associated molecular pattern and aggravates acute liver injury via promoting inflammation. Free Radic Biol Med 137:24-36 (2019). Read more (PubMed: 30991142) »