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Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric)

Highly Sensitive; Includes Paraoxonase Positive Control
Catalog #: K999
$525.00

Product Details

Cat # +Size K999-100
Size 100 assays
Detection Method Fluorescence (Ex/Em 368/460 nm)
Species Reactivity Eukaryotes
Applications • Rapid assessment of native/recombinant Paraoxonase
- Paraoxonase activity in mammalian serum and plasm
Features & Benefits • Simple, convenient, highly sensitive · Fluorescent assay enables easy determination of PON1 activity in a variety of biological samples
• The substrate shows low background and a high signal-to-noise ratio
• Kit includes Paraoxonase inhibitor, 2-hydroxyquinoline, and a stable, Paraoxonase as positive control
Kit Components • Paraoxonase Assay Buffer
• Fluorescence Standard • PON1 Inhibitor (2-hydroxyquinoline)
• PON1 Substrate
• Paraoxonase Positive Control
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.

Details

Paraoxonase 1 (PON1, EC 3.1.8.1) is a 43 kDa enzyme synthesized in the liver and released into the bloodstream, where it associates with high-density lipoprotein (HDL) particles in serum. PON1 is a promiscuous enzyme with broad-spectrum hydrolase activity that was originally named for its ability to hydrolyze highly toxic organophosphorus compounds such as the insecticide paraoxon and various nerve agents used as chemical weapons. The enzyme has subsequently been demonstrated to catalyze hydrolysis of lipid hydroperoxides and lactones. PON1 helps protect serum HDL and LDL particles against lipid peroxidation and inhibits N-homocysteinylation of LDL-associated proteins by hydrolyzing the highly reactive pro-oxidant homocysteine thiolactone. Accumulation of lipid peroxides and homocysteinylated proteins triggers arterial inflammation and eventually leads to atherosclerosis, ischemic stroke and myocardial infarction. PON1 activity is considered to be a clinical biomarker of hepatic and systemic oxidative stress. Measurement of serum PON1 activity has been proposed as a potential test for evaluation of liver function and risk of cardiovascular disease. BioVision’s Paraoxonase 1 Activity Assay Kit enables rapid measurement of PON1 activity, utilizing a fluorogenic substrate that is converted into a highly fluorescent product (Ex/Em = 368/460 nm). This ensures dramatically greater sensitivity than UV or colorimetric assays and eliminates the need for dangerous toxic substrates. A selective PON1 inhibitor is provided for verification of PON1 specific activity. The assay is simple to perform, high-throughput adaptable and can detect a minimum of 250 μU paraoxonase activity with a sample volume of 5 μl


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I have run my serum samples and my values are 10,000 fold higher than what is found in literature. Is there any explanation as to why this is happening? Do you have any data regarding Vmax for more common substrates like phenyl acetate relative to your proprietary fluorescently labelled substrate?
The substrate used in the traditional PON1 activity assay, namely Phenyl acetate has much lower affinity and has much faster Vmax for the PON1 enzyme. The PON1 enzyme has 3 activities, organic phosphate ester activity, general esterase activity (for phenyl acetate substrate) and hydrolysis of homocysteinethiolactone. We are measuring the organophosphatase activity of the enzyme (cleaves organic phosphate esters). The substrate we use is more selective for PON1 than other commonly used substrates such as Phenyl Acetate. Therefore, you will see higher activity.