The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml.
Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).
Use a concentration of 1 µg/ml.
Use a concentration of 5 µg/ml.
FunctionInvolved in redox regulation of the cell. Can reduce H(2)O(2) and short chain organic, fatty acid, and phospholipid hydroperoxides. May play a role in the regulation of phospholipid turnover as well as in protection against oxidative injury.
Sequence similaritiesBelongs to the ahpC/TSA family. Rehydrin subfamily. Contains 1 thioredoxin domain.
Cellular localizationCytoplasm. Lysosome. Cytoplasmic vesicle. Also found in lung secretory organelles.
ICC/IF image of ab73350 stained HeLa cells. The cells were 4% PFA fixed (10 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 (ab73350, 1µg/ml) 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. This antibody also gave a positive result in 100% methanol fixed (10 min) HeLa, HepG2, Hek293 and MCF7 cells at 1µg/ml.
IHC image of Peroxiredoxin 6 staining in Human Hippocampus FFPE section, performed on a 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 ab73350, 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
Western blot - Anti-Peroxiredoxin 6 antibody (ab73350)
Anti-Peroxiredoxin 6 antibody (ab73350) at 1 µg/ml + Recombinant human Peroxiredoxin 6 protein (ab87631) at 0.01 µg
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique
Peroxiredoxin 6 was immunoprecipitated using 0.5mg Rat Testis tissue lysate, 5µg of Rabbit polyclonal to Peroxiredoxin 6 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Rat Testis tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab73350. Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution. Band: 25KDa: Peroxiredoxin 6
References for Anti-Peroxiredoxin 6 antibody (ab73350)
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