Active Paraoxonase 3 (PON3)

Serum paraoxonase/lactonase 3

ACTIVITY TEST

Paraoxonase 3 (PON3) is secreted into the bloodstream and associates with high-density lipoprotein (HDL). PON3 also rapidly hydrolyzes lactones and can inhibit the oxidation of low-density lipoprotein, a function that is believed to slow the initiation and progression of atherosclerosis. Besides, Paraoxonase 1 (PON1) has been identified as an interactor of PON3, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PON3 and recombinant human PON1. Briefly, PON3 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to PON1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PON3 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of of PON3 and PON1 was shown in Figure 1, and this effect was in a dose dependent manner.

USAGE

Reconstitute in 20mM Tris, 150mM NaCl (pH8.0) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Journal of Pharmaceutical and Biomedical Analysis Role of esterase mediated hydrolysis of simvastatin in human and rat blood and its impact on pharmacokinetic profiles of simvastatin and its active metabolite in rat Pubmed: 30776567
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