Anti-2 Cys Peroxiredoxin antibody [6E5] (ab16765)

Overview

  • Product nameAnti-2 Cys Peroxiredoxin antibody [6E5]
  • Description
    Mouse monoclonal [6E5] to 2 Cys Peroxiredoxin
  • Tested applicationsSuitable for: ELISA, WB, Flow Cyt, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Rat, Human, Escherichia coli
  • Immunogen

    Recombinant full length protein (Human).

  • Positive control
    • HeLa whole cell lysate. IF/ICC: HCT116 cell line.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • 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
  • Database links
  • Alternative names
    • Antioxidant enzyme antibody
    • Peroxiredoxin 1 antibody
    • Peroxiredoxin 2 antibody
    • Peroxiredoxin 3 antibody
    • Peroxiredoxin 4 antibody
    • Peroxiredoxin 5 antibody
    • Peroxiredoxin 5 mitochondrial antibody
    • PLP antibody
    • PMP20 antibody
    • PRDX1 antibody
    • PRDX2 antibody
    • PRDX3 antibody
    • PRDX4 antibody
    • PRDX5 antibody
    • PRX V antibody
    • Thioredoxin reductase antibody
    see all

Anti-2 Cys Peroxiredoxin antibody [6E5] images

  • 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



  • Predicted band size : 24 kDa

    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.

References for Anti-2 Cys Peroxiredoxin antibody [6E5] (ab16765)

This product has been referenced in:
  • Nardozza AP  et al. Reactive oxygen species and epidermal growth factor are antagonistic cues controlling SHP-2 dimerization. Mol Cell Biol 32:1998-2009 (2012). WB ; Human . Read more (PubMed: 22411627) »
  • O'Neill JS & Reddy AB Circadian clocks in human red blood cells. Nature 469:498-503 (2011). WB ; Human . Read more (PubMed: 21270888) »

See all 6 Publications for this product

Product Wall

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (4-12% NuPAGE Bis-Tris gel, MES buffer, antioxidant in top tank)
Sample Saccharomyces cerevisiae Cell lysate - whole cell (Whole cell extract prepared using TCA pptn)
Specification Whole cell extract prepared using TCA pptn
Treatment Inc'd for 15' with 1mM H2O2, or no peroxide
Blocking step 0.5% w/w BSA, 0.5% non-fat dried milk (Marvel) in Tris buffered saline/0.05% Tween-20 (TBST). as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.5% · Temperature: ~20°C
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Verified customer

Submitted Oct 29 2014

Thank you for the inquiry. We produce some of our antibodies in-house and others are sourced from suppliers whose products we believe will be of interest to our customers. This product falls into the latter category. The identity of our suppliers remai...

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Thank you for your enquiry. I understand your customers difficulty. I am in touch with the source of ab16765 to determine the isoform used to raise this antibody. I can tell you that this antibody recognizes PrxI, PrxII, PrxIII and PrxIV by western...

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