Overview

  • Product name
    Anti-Peroxiredoxin 1/PAG antibody
    See all Peroxiredoxin 1/PAG primary antibodies
  • Description
    Rabbit polyclonal to Peroxiredoxin 1/PAG
  • Host species
    Rabbit
  • Specificity
    This 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 applications
    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, African green monkey
    Predicted to work with: Cow, Chinese hamster
  • Immunogen

    Synthetic peptide corresponding to Human Peroxiredoxin 1/PAG aa 103-114.
    Sequence:

    LVSDPKRTIAQD

  • Positive control
    • 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.

Properties

Applications

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.

Application Abreviews Notes
ICC/IF 1/200.
IHC-P Use at an assay dependent concentration.
WB 1/1000. Detects a band of approximately 20 kDa (predicted molecular weight: 22 kDa).

Target

  • Function
    Involved 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 similarities
    Belongs to the ahpC/TSA family.
    Contains 1 thioredoxin domain.
  • Post-translational
    modifications
    Phosphorylated on Thr-90 during the M-phase, which leads to a more than 80% decrease in enzymatic activity.
  • Cellular localization
    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • Heme binding 23 kDa protein antibody
    • MSP23 antibody
    • Natural killer cell-enhancing factor A antibody
    • NKEF A antibody
    • NKEF-A antibody
    • NKEFA antibody
    • OSF3 antibody
    • Osteoblast specific factor 3 antibody
    • PAG antibody
    • Paga antibody
    • PAGB antibody
    • Peroxiredoxin 1 antibody
    • Peroxiredoxin-1 antibody
    • PRDX1 antibody
    • PRDX1_HUMAN antibody
    • Proliferation associated gene A antibody
    • Proliferation-associated gene protein antibody
    • PRX1 antibody
    • PrxI antibody
    • TDPX2 antibody
    • Thioredoxin peroxidase 2 antibody
    • Thioredoxin-dependent peroxide reductase 2 antibody
    see all

Images

  • 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


    Secondary
    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.

References

This product has been referenced in:
  • Li HX  et al. Peroxiredoxin 1 promoted tumor metastasis and angiogenesis in colorectal cancer. Pathol Res Pract 214:655-660 (2018). Read more (PubMed: 29673884) »
  • Iqbal J  et al. Selenium positively affects the proteome of 3 × Tg-AD mice cortex by altering the expression of various key proteins: unveiling the mechanistic role of selenium in AD prevention. J Neurosci Res 96:1798-1815 (2018). Read more (PubMed: 30054946) »
See all 18 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
50 µg
Specification
Skeletal muscle
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 10 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (15%)
Loading amount
50 µg
Specification
Skeletal muscle
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 10 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkats)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20000 cells
Specification
Jurkats
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Dr. Tejpal Gill

Verified customer

Submitted Jan 13 2014

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (brain)
Permeabilization
Yes - triton 0.1%
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Fixative
Paraformaldehyde

Dr. Paul Fraser

Verified customer

Submitted Dec 16 2013

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxxxx. To check the status of the order please contact our Customer Service team and reference this number. Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know. I wish you the best of luck with your research.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle (AT))
Gel Running Conditions
Reduced Denaturing (12% gel)
Loading amount
50 µg
Specification
Skeletal muscle (AT)
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Dr. G.K Sakellariou

Verified customer

Submitted Sep 28 2012

Answer

Merci de nous avoir contactés.

Nous sommes désolés d'apprendre que le produit ab15571 que vous avez reçu ne fonctionne pas comme attendu.
Le numéro de commande de remplacement gratuitavec leproduit ab41906 est *****. Vous recevrez prochainement un mail de confirmation comprenant les détails d'expédition.

N'hésitez pas à nous contacter lors d'une prochaine occasion.

Read More

Answer

Thank you for your telephone enquiry yesterday.

As requested, I am forwarding a list of suitable secondary antibodies for use with ab15571 and ab6276. These have been tested and guaranteed in ICC-IF. I have selected some FITC and PE conjugated antibodies. I can recommend to review the online datasheets for further information.

For the double staining, you will need to select an FITC conjugated secondary for one of the primaries, and a PE conjugated secondary for the other. I have provided a selection of both below, so you can make your own choice.

Secondaries for use with ab6276 mouse IgG1:

FITC conjugated:

ab97239 Goat anti-Mouse IgG1 heavy chain (FITC) secondary antibody
Tested in: Flow Cyt, ICC/IF, IHC-P
https://www.abcam.com/index.html?datasheet=97239 (or use the following: https://www.abcam.com/index.html?datasheet=97239).

ab98692 Goat polyclonal Secondary Antibody to Mouse IgG1 - heavy chain (FITC), pre-adsorbed
Tested in: Flow Cyt, ICC/IF, IHC-P
https://www.abcam.com/index.html?datasheet=98692 (or use the following: https://www.abcam.com/index.html?datasheet=98692).

PE conjugated:

ab99605 Rat monoclonal [SB77e] Secondary Antibody to Mouse IgG1 (PE), pre-adsorbed
Tested in: ELISA, Flow Cyt, ICC/IF
https://www.abcam.com/index.html?datasheet=99605 (or use the following: https://www.abcam.com/index.html?datasheet=99605).


Secondaries for use with ab15571 Rabbit polyclonal:

FITC conjugated:

ab97050 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (FITC)
Tested in: Flow Cyt, ICC/IF, IHC-P
https://www.abcam.com/index.html?datasheet=97050 (or use the following: https://www.abcam.com/index.html?datasheet=97050).

ab97199 Goat polyclonal Secondary Antibody to Rabbit IgG - Fc (FITC)
Tested in: Flow Cyt, ICC/IF, IHC-P
https://www.abcam.com/index.html?datasheet=97199 (or use the following: https://www.abcam.com/index.html?datasheet=97199).

PE conjugated:

ab7007 Donkey F(ab')2 polyclonal Secondary Antibody to Rabbit IgG - H&L (PE), pre-adsorbed
Tested in: Flow Cyt, ICC/IF, IHC-P, IM
https://www.abcam.com/index.html?datasheet=7007 (or use the following: https://www.abcam.com/index.html?datasheet=7007).

ab50677
Goat anti-Rabbit IgG H&L (PE) secondary antibody
Tested in: ICC-IF.
https://www.abcam.com/index.html?datasheet=50677 (or use the following: https://www.abcam.com/index.html?datasheet=50677).

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Answer

We have previously been in contact regarding ab15571, which was not providing satisfactory results.

We provided some troubleshooting tips for the protocol, and I am writing to inquire if these suggestions have helped to improve the results? I would be pleased to receive an update on how these experiments are progressing.

If the antibody is still not working as stated on the datasheet, and the item was purchased within 6 months of contacting us, we would be happy to replace or refund the antibody.

I wish you the best of luck with your research and I look forward to hearing from you. Please do not hesitate to contact us if you have any further questions.

Read More

Answer

Thank you for your email.

I am sorry without the positive control it will be hard to determine whether the no band problem is due to antibody, protocol or target being not expressed in cell line you have been using.

We have lysates of these cell lines in catalogue. If you are interested I can send it to you at discounted price.
Human protein Atlas shows this protein is expressed in quite a range of tissue so please click the following link to explore more positive control

http://www.proteinatlas.org/ENSG00000117450/normal

Finally I would like to ask if you have found any publication which shows the protein expression in lymphocytes and HdhQ7/Q7 cells.

I hope this information will be helpful.

Read More

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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