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Anti-Peroxiredoxin 3抗体(ab73349)

概述

  • 产品名称Anti-Peroxiredoxin 3抗体
    参阅全部 Peroxiredoxin 3 一抗
  • 描述
    兔多克隆抗体to Peroxiredoxin 3
  • 经测试应用适用于: IHC-P, ICC/IF, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
    预测可用于: Chicken, Cow, Xenopus laevis, Chimpanzee, Zebrafish, Orangutan
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 200 to the C-terminus of Human Peroxiredoxin 3.

    (Peptide available as ab85815.)

  • 阳性对照
    • This antibody gave a positive signal in the following lysates: Human, Mouse and Rat Liver, Rat Ovary.

性能

应用

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.

应用 Ab评论 说明
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).

靶标

Anti-Peroxiredoxin 3 antibody 图像



  • Predicted band size : 28 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Peroxiredoxin 3 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 when Peroxiredoxin 3 knockout samples were used. Wild-type and Peroxiredoxin 3 knockout samples were subjected to SDS-PAGE. ab73349 and ab8245 (loading control to GAPDH) were diluted at 1 μg/ml and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-Peroxiredoxin 3 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
    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 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.

Anti-Peroxiredoxin 3 antibody (ab73349)参考文献

This product has been referenced in:
  • Bhatt NM  et al. Restoring redox balance enhances contractility in heart trabeculae from type 2 diabetic rats exposed to high glucose. Am J Physiol Heart Circ Physiol 308:H291-302 (2015). WB ; Rat . Read more (PubMed: 25485897) »
  • Cao Z  et al. Lack of an efficient endoplasmic reticulum-localized recycling system protects peroxiredoxin IV from hyperoxidation. J Biol Chem 289:5490-8 (2014). WB ; Human . Read more (PubMed: 24403061) »

See all 2 Publications for this product

Product Wall

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
Username

Abcam user community

Verified customer

提交于 Sep 23 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Sep 19 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Rat Cell lysate - other (Ovary)
Specification Ovary
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Sep 09 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 75 µg
Gel Running Conditions Reduced Denaturing (12.5)
Sample Rat Cell lysate - whole cell (Ovarian granulosa cells)
Specification Ovarian granulosa cells
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Dr. Sakhila Banu

Verified customer

提交于 Aug 12 2013

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 ...

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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 diffe...

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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 d...

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

Dr. Sophie Pezet

Verified customer

提交于 Sep 08 2010

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