Serum paraoxonase/arylesterase 1 (PON1) also known as aromatic esterase 1 or serum aryldialkylphosphatase 1 is an enzyme that in humans is encoded by the PON1 gene. The PON gene was mapped to chromosome 7q21-q22, and the mouse Pon1 gene was mapped to the proximal end of chromosome 6. PON1 is responsible for hydrolysing organophosphate pesticides and nerve gasses. Polymorphisms in the PON1 gene significantly affect the catalytic ability of the enzyme. PON1 (paraoxonase 1) is also a major anti-atherosclerotic component of high-density lipoprotein (HDL). Sample Type: Cell culture supernates, serum, plasma (Heparin). Intended Use: Sandwich High Sensitivity BioAssay(TM) ELISA kit for Quantitative Detection of Mouse PON1/Paraoxonase 1. Sensitivity: <10pg/ml Range: 156pg/ml-10,000pg/ml Specificity: Recognizes natural and recombinant Mouse PON1/Paraoxonase 1. There is no detectable cross-reactivity with other relevant proteins. Test Principle: This Mouse PON1/Paraoxonase 1 BioAssay(TM) ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A a monoclonal antibody from rat specific for PON1/Paraoxonase 1 has been precoated onto 96-well plates. Standards (Expression system for standard: E.Coli, Immunogen sequence: A30-L355) and test samples are added to the wells, a biotinylated detection a biotinylated polyclonal antibody from goat specific for PON1/Paraoxonase 1 is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Mouse PON1/Paraoxonase 1 amount of sample captured in plate. Kit Components: 96-well plate precoated with anti-Mouse Pon1 antibody, 1x plate Lyophilized recombinant Mouse Pon1 standard, 2x10ng/tube Biotinylated anti-Mouse Pon1 antibody, 1x130ul (dilution 1:100) Avidin-Biotin-Peroxidase Complex (ABC), 1x130ul (dilution 1:100) Sample diluent buffer, 1x30ml Antibody diluent buffer, 1x12ml ABC diluent buffer, 1x12ml TMB color developing agent, 1x10ml TMB stop solution, 1x10ml PBS washing buffer Powder for 1000ml, 1x buffer Storage and Stability: Store powder at 4C liquid at -20C. Store other components at 4C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
