概述

  • 产品名称
  • 描述
    兔多克隆抗体to PON3
  • 经测试应用
    适用于: ICC/IF, IHC-P, WBmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human PON3.

    (Peptide available as ab42321.)

  • 阳性对照
    • WB: Human skeletal muscle, mouse liver, rat liver and rat skeletal muscle tissue lysates. ICC/IF: HepG2 cells.

性能

应用

Our Abpromise guarantee covers the use of ab42322 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
ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 40 kDa).

靶标

图片

  • ICC/IF image of ab42322 stained HepG2 cells. The cells were 4% formaldehyde 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 (ab42322, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit 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.

  • All lanes : Anti-PON3 antibody (ab42322) at 1 µg/ml

    Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 2 : Liver (Mouse) Tissue Lysate - normal tissue
    Lane 3 : Liver (Rat) Tissue Lysate
    Lane 4 : Skeletal Muscle (Rat) Tissue Lysate - normal tissue (ab29376)

    Lysates/proteins at 10 µg per lane.

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

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 43 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 49 kDa. We are unsure as to the identity of these extra bands.
  • IHC image of ab42322 staining PON3 in Human liver 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 ab42322, 1µg/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.

    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.

文献

This product has been referenced in:
  • Pei JF  et al. Human paraoxonase gene cluster overexpression alleviates angiotensin II-induced cardiac hypertrophy in mice. Sci China Life Sci 59:1115-1122 (2016). Read more (PubMed: 27578362) »
  • Birkus G  et al. Intracellular Activation of Tenofovir Alafenamide and the Effect of Viral and Host Protease Inhibitors. Antimicrob Agents Chemother 60:316-22 (2015). Read more (PubMed: 26503655) »

See all 3 Publications for this product

客户评价及客户问答

The PON3 antibody that I would like to send to you free-of-charge is ab108964. This is a rabbit monoclonal that has given good western blot results for a variety of human tissue and cell lysates. However, it is not suitable for IHC or ICC.

C...

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Thank you for your email and the additional information. This is very helpful.

I would suggest that if no protease inhibitors were used, to include them in your lysis buffer freshly, before lysing the cells (e.g. ab65621 protease Inhibitor c...

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Thank you for bringing this to our attention.

I am sorry to read that you are having difficulty obtaining a western blot signal. I may be able to make some recommendations but I will need a few more details about your samples and protocol. <...

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

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