Product nameAnti-2 Cys Peroxiredoxin antibody [6E5]
DescriptionMouse monoclonal [6E5] to 2 Cys Peroxiredoxin
Tested applicationsSuitable for: ELISA, WB, Flow Cyt, IHC-P, ICC/IFmore details
Species reactivityReacts with: Rat, Human, Escherichia coli
Recombinant full length protein (Human).
- HeLa whole cell lysate. IF/ICC: HCT116 cell line.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferPreservative: 0.03% Sodium azide
Constituents: HEPES, 50% Glycerol, 0.87% Sodium chloride, 0.01% BSA
Concentration information loading...
Purification notesPurified from ascites.
Light chain typekappa
Our Abpromise guarantee covers the use of ab16765 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ELISA||Use at an assay dependent concentration.|
|WB||1/1000. Predicted molecular weight: 24 kDa.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||Use a concentration of 10 µg/ml.|
RelevancePeroxiredoxin (Prx) is a growing peroxidase family whose mammalian members are involved in cell proliferation, differentiation, and apoptosis. There are many isoforms(about 50 proteins), known on the basis of amino acid sequence homology, particularly the amino-terminal region containing an active site cysteine residue. The thiol-specific antioxidant activity is distributed throughout all the kingdoms. Among them, mammalian Prx consists of 6 different members grouped into typical 2 Cys, atypical 2 Cys Prx, and 1 Cys Prx. Except PrxVI belonging to the 1 Cys Prx subgroup, the other five 2 Cys Prx isotypes share the thioredoxin dependent peroxidase(TPx) activity utilizing thioredoxin, thioredoxin reductase, and NADPH as a reducing system. Mammalian Prxs are 20–30 kilodaltons in molecular size and vary in subcellular localization:PrxI, II and VI in cytosol, PrxIII in mitochondria, PrxIV in ER and secretion, PrxV showing complicated distribution including peroxisome, mitochondria and cytosol.
Cellular localizationCytoplasmic and Mitochondrial
- Antioxidant enzyme antibody
- Peroxiredoxin 1 antibody
- Peroxiredoxin 2 antibody
ICC/IF image of ab16765 stained HCT116 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 (ab16765, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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
Western blot analysis using ab16765 at 1/1000 dilution using HeLa cell lysates:
Lane 1: Recombinant human PrxI proetein
Lane 2: Recombinant human PrxII protein
Lane 3: Recombinant human PrxIII protein
Lane 4: Recombinant human PrxIV protein (w/o secretion leader sequence)
Lane 5: HeLa cell lysates.Western blot analysis using ab16765 at 1/1000 dilution using HeLa cell lysates: Lane 1: Recombinant human PrxI proetein Lane 2: Recombinant human PrxII protein Lane 3: Recombinant human PrxIII protein Lane 4: Recombinant human PrxIV protein (w/o secretion leader sequence) Lane 5: HeLa cell lysates.
Overlay histogram showing HeLa cells stained with ab16765 (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. The cells were then incubated with the antibody (ab16765, 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 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.
This product has been referenced in:
- Zhang ZH et al. The effect of subarachnoid erythrocyte lysate on brain injury: a preliminary study. Biosci Rep 36:N/A (2016). WB . Read more (PubMed: 27279653) »
- Milev NB et al. Analysis of the redox oscillations in the circadian clockwork. Methods Enzymol 552:185-210 (2015). Read more (PubMed: 25707278) »