Product datasheet
Anti-2 Cys Peroxiredoxin antibody [6E5] ab16765
1 Abreviews 6 References 3 Images
Overview
Product name
Anti-2 Cys Peroxiredoxin antibody [6E5]
Description
Mouse monoclonal [6E5] to 2 Cys Peroxiredoxin
Tested applications
ELISA, WB, Flow Cyt, IHC-P, ICC/IF
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
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
Preservative: 0.03% Sodium Azide
Constituents: 50% Glycerol, 0.01% BSA, HEPES, 0.15M Sodium chloride
Purification notes
Purified by ammonium precipitation
Clonality
Monoclonal
Clone number
6E5
Isotype
IgG1
Light chain type
kappa
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.
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Application
Abreviews
Notes
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
Relevance
Peroxiredoxin (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 localization
Cytoplasmic and Mitochondrial
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
Immunocytochemistry/ Immunofluorescence -
anti-mouse IgG (H+L) used at a 1/250 dilution
Anti-2 Cys Peroxiredoxin antibody [6E5]
for 1h. Alexa Fluor® 594 WGA was used to
(ab16765)
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
2
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
Western blot - 2 Cys Peroxiredoxin antibody
[6E5] (ab16765)
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.
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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
Flow Cytometry-Anti-2 Cys Peroxiredoxin
with the antibody (ab16765, 1µg/1x106 cells)
antibody [6E5](ab16765)
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.
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