Overview

  • Product nameAnti-PON2 antibodySee all PON2 primary antibodies ...
  • Description
    Rabbit polyclonal to PON2
  • Tested applicationsIHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Cow, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human PON2.

    (Peptide available as ab41542.)

  • Positive control
    • This antibody gave a positive signal in the following Human whole cell lysates: Caco 2; HeLa - Hydroxyurea Treated (48hr, 1µM). HeLa cell line in IF.

Properties

Applications

Our Abpromise guarantee covers the use of ab40969 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 1/2000.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).Can be blocked with Human PON2 peptide (ab41542).
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-PON2 antibody images

  • Immunohistochemistical detection of PON2 antibody (ab40969) on  formaldehyde fixed paraffin-embedded mosue kidney and lung sectionsAntigen retrieval step: Heat mediated. Buffer: Citric acid pH6. Permeabilization: None. Blocking step: 1% BSA for 10 mins @ 21°C. Primary antibody incubated at 1/2000 for 2 hours @ 21°C in TBS/BSA/azide. Secondary antibody: anti Rabbit IgG Conjugated to Biotin (1/200). Patterns of immunostaining are produced that appear to be very similar to those seen on the HPA website, for these tissues. In the submitted composite image, both tissues show intense labelling. In the upper image of kidney cortico-medullary junction, apical membrane positivity is observed in proximal/distal tubules. In the same tubules globular intra-cytoplasmic positivity is also observed. In the lower image of lung, positivity is evident in the simple columnar epithelium of the bronchioles and also in what appear to be alveolar macrophages. This antibody should be diluted further, under the conditions used.

    See Abreview

  • All lanes : Anti-PON2 antibody (ab40969) at 1 µg/ml

    Lane 1 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 2 : Hela Whole Cell Lysate - Hydroxyurea Treated (48hr, 1uM)

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 40 kDa
    Observed band size : 40 kDa
    Additional bands at : 45 kDa (possible glycosylated form),55 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 150 seconds
  • Anti-PON2 antibody (ab40969) at 1 µg/ml + Human PON2 full length protein (ab53379) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 30 seconds
  • ICC/IF image of ab40969 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine for 1h to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab40969, 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.

References for Anti-PON2 antibody (ab40969)

This product has been referenced in:
  • Giordano G  et al. Gender differences in brain susceptibility to oxidative stress are mediated by levels of paraoxonase-2 expression. Free Radic Biol Med 58:98-108 (2013). WB ; Mouse, Human . Read more (PubMed: 23376469) »

See 1 Publication for this product

Product Wall

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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Thank you for contacting us.

We sell this product at a 100ul size. How many western blots you will be able to perform with that depends upon a number of factors including the concentration that is needed for your optimized protocol and the...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Kidney and lung)
Specification Kidney and lung
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Nov 02 2010

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