Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric) (BN01156)

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BN01156
  • Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric) (BN01156)
  • Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric) (BN01156)
  • Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric) (BN01156)
€708

Description

ELISA Kit Technical ManualMSDS

Paraoxonase 1 (PON1) Activity Assay Kit (Fluorometric)

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. Assay Genie'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 paraoxonase activity with a sample volume of 5

Figure: (a) Standard curve of PON1 Substrate metabolite fluorescence. One mole of fluorescence standard corresponds to the metabolism of one mole of PON1 Substrate. (b) Reaction kinetics of PON1 Substrate metabolism in donor-pooled human serum (5 µl), donor-pooled human plasma (5 µl) and rabbit serum (2.5 µl) in the presence and absence of 200 µM of the selective PON1 inhibitor 2-hydroxyquinoline (no inhibitor conditions contained 0.4% DMSO as a solvent control) (c) Quantification of PON1 activity in serum/plasma samples (mean ± SEM of four independent replicates). Assays were performed according to the kit protocol.

Key Information Description

Product SKU

BN01156

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 and 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.

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