Overview

  • Product name

    Mouse PON3 ELISA Kit
    See all PON3 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Plasma 5 4%
    Inter-assay
    Sample n Mean SD CV%
    Plasma 3 4.1%
  • Sample type

    Cell culture supernatant, Serum, Plasma, Cell culture extracts, Tissue Extracts
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    5.36 pg/ml
  • Range

    0.23 ng/ml - 15 ng/ml
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Cell culture supernatant 97 93% - 101%
    Serum 110 108% - 111%
    Cell culture extracts 102 101% - 104%
    Tissue Extracts 112 108% - 115%
    EDTA Plasma 109 107% - 111%
    Citrate Plasma 110 107% - 112%
    Fetal Bovine Serum 99 98% - 99%

  • Assay time

    1h 30m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse
  • Product overview

    PON3 in vitro SimpleStep ELISA® (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of PON3 protein in serum, plasmas, cell culture supernatants, and cell and tissue extracts.


    The SimpleStep ELISA® employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material.  TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Notes

    Serum paraoxonase/lactonase 3 (PON3) is a calcium binding 354 amino acid protein. PON3 possesses hydrolytic catalytic activity against aromatic and 5- or 6- member ring lactones. PON3 is a member of the paraoxonase family consisting of PON1 and PON, which are involved in modulating oxidative stress and possess anti-atherogenic properties. PON3 is a secreted protein that associates with high-density lipoprotein (HDL).

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

Associated products

Applications

Our Abpromise guarantee covers the use of ab211653 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Interpolated concentrations of spiked recombinant PON3 in pooled normal mouse serum, plasma (citrate), and cell culture media (RPMI 1640 + 10% fetal bovine serum) samples, and native PON3 in plasma (EDTA) samples.  The concentrations of PON3 were measured in duplicate, interpolated from the PON3 standard curves and corrected for sample dilution. Undiluted samples are as follows:  serum 10%, plasma (citrate) 10%, plasma (EDTA) 20% and cell culture media 32.5%.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). 

  • The concentrations of PON3 were measured in duplicate and interpolated from the PON3 standard curve and corrected for sample dilution.  The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Dilutions of pooled normal mouse serum and plasma within the linear range were measured in duplicate, their background-subtracted values interpolated off of the mouse PON3 standard curve, and corrected for dilution back to neat. The mean PON3 concentration was determined to be 6.7 ng/mL in serum, 4.5 ng/mL in plasma (citrate), 17.8 ng/mL in plasma (EDTA), and 6 ng/mL in RAW 264.7 cell culture supernatant.

  • Dilutions of cell and tissue extracts within the linear range were measured in duplicate, their background-subtracted values interpolated off of the mouse PON3 standard curve, and corrected for dilution back to a 1000 µg extract load. The mean PON3 concentration was determined to be 71.9 ng/mg in liver extract and 669 ng/mg in RAW 264.7 cell extract.

Protocols

References

ab211653 has not yet been referenced specifically in any publications.

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