Overview

  • Product name

    Anti-Peroxiredoxin 3/PRDX3 antibody
    See all Peroxiredoxin 3/PRDX3 primary antibodies
  • Description

    Rabbit polyclonal to Peroxiredoxin 3/PRDX3
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Chicken, Cow, Xenopus laevis, Chimpanzee, Zebrafish, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human Peroxiredoxin 3/PRDX3 aa 200 to the C-terminus conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab85815)

  • Positive control

    • Recombinant human Peroxiredoxin 3/PRDX3 protein (ab85559) can be used as a positive control in WB. This antibody gave a positive signal in the following lysates: Human, Mouse and Rat Liver, Rat Ovary.
  • General notes

     This product was previously labelled as Peroxiredoxin 3

     

Properties

Applications

Our Abpromise guarantee covers the use of ab73349 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
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 28 kDa).

Target

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Peroxiredoxin 3/PRDX3 knockout HAP1 cell lysate (20 µg)
    Lane 3: MCF7 cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab73349 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab73349 was shown to specifically react with Peroxiredoxin 3/PRDX3 in wild-type HAP1 cells. No band was observed when Peroxiredoxin 3/PRDX3 knockout samples were examined. Wild-type and Peroxiredoxin 3/PRDX3 knockout samples were subjected to SDS-PAGE. ab73349 and ab8245 (loading control to GAPDH) were diluted at 1μg/ml and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-Peroxiredoxin 3/PRDX3 antibody (ab73349) at 1 µg/ml

    Lane 1 : Human liver tissue lysate - total protein (ab29889)
    Lane 2 : Liver (Rat) Tissue Lysate
    Lane 3 : Ovary (Rat) Tissue Lysate
    Lane 4 : Liver (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 28 kDa
    Observed band size: 23 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 45 kDa, 52 kDa. We are unsure as to the identity of these extra bands.

  • ICC/IF image of ab73349 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 (ab73349, 5µg/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 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml.
  • IHC image of Peroxiredoxin 3/PRDX3 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 ab73349, 5µg/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.

References

This product has been referenced in:

  • Booty LM  et al. Selective Disruption of Mitochondrial Thiol Redox State in Cells and In Vivo. Cell Chem Biol 26:449-461.e8 (2019). Read more (PubMed: 30713096) »
  • Brown LA  et al. Late life maintenance and enhancement of functional exercise capacity in low and high responding rats after low intensity treadmill training. Exp Gerontol 125:110657 (2019). Read more (PubMed: 31306740) »
See all 17 Publications for this product

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Baboon Tissue sections (skeletal muscle)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10 mM Sodium Citrate (pH 6.0)
Permeabilization
No
Specification
skeletal muscle
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Dr. Soon-Ok Kim

Verified customer

Submitted Oct 02 2017

Application
Western blot
Sample
Cow Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (Used any kD precast gel)
Loading amount
25 µg
Specification
Liver
Blocking step
Milk as blocking agent for 40 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 23 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
Antigen retrieval step
Enzymatic
Sample
Rat Tissue sections (GONAD)
Specification
GONAD
Permeabilization
No
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Sep 19 2013

Application
Western blot
Sample
Rat Cell lysate - other (Ovary)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
75 µg
Specification
Ovary
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 09 2013

Application
Western blot
Sample
Rat Cell lysate - whole cell (Ovarian granulosa cells)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
75 µg
Specification
Ovarian granulosa cells
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Sakhila Banu

Verified customer

Submitted Aug 13 2013

Answer

According to our records, ab16751 was proving difficult to use inWB and we were in contact in order to help resolve the issue.

Looking at our correspondence, it appears that we are awaiting more details in order to help us better understand the difficulties experienced. If the requested information has already been sent, it appears that it did not reach our Scientific Support team and we apologize for this inconvenience. In this case we would like to ask for the information again so that we can reach a resolution.

If the issue has already been settled, please let us know so that we can be assured that the problem has been solved to your satisfaction and update our records.

We wish you the best of luck with your research and look forward to a reply.

Read More

Answer

Thank you for contacting us. The antibody should be detecting a band around 27 kDa. I would suggest searching and literature and comparing with other people findings. The NCBI Ref seq actually gives info of two isoforms which are slightly different to each other. It may be possible that the antibody is detecting a different isoform than canonical. It is just a thought; you may need to check the literature or running further tests would also be an option. I am sorry I can’t be more specific as I don’t have information of protocol you have been using. Please check the following links: http://www.ncbi.nlm.nih.gov/protein/NP_006784.1 http://www.uniprot.org/uniprot/P30048 http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jobId=clustalw2-I20111109-155629-0237-39221683-pg I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

Read More

Answer

Thank you for contacting us. I am sorry to hearing about the confusion. We never send wrong products to our customers so if you have ordered antibody ab16751 then the antibody in the vial will be Anti-Peroxiredoxin 3 antibody [12B] (ab16751).  We do send different products in cases where the products are out of stock but not without confirmation from our customers. We always contact them by phone or by email for their choice of waiting or selecting another product from catalogue. The ab16751 was in stock when you ordered it so there is not room for error from our end. I also have few questions which will help me to understand the problem: Ab16751 is a Mouse monoclonal and ab73349 is a Rabbit polyclonal. Could you specify which secondary antibody you have used; was it anti mouse or anti rabbit. The extra bands between 50-60 kDa do not mean that the antibody is different in the vial. These bands could be due to different reasons e.g. high conc. of primary or secondary antibody, loading high conc. of lysates, not using protease inhibitors in lysis buffer etc. We have observed 2 extra bands in case of ab73349 with Rat liver lysates; are you using the same lysates? Finally I would suggest troubleshooting the experiment. I can also help you with troubleshooting however I would need more information about the protocol used. Please fill in the attached questionnaire; I will then check the protocol for possible optimizations. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

Read More
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain - Hippocampus)
Specification
Brain - Hippocampus
Fixative
Paraformaldehyde

Dr. Sophie Pezet

Verified customer

Submitted Sep 08 2010

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