Alkaline Phosphatase Enzyme

β-Galactosidase Enzyme

β-Glucuronidase Enzyme

β-Glucosidase Enzyme

Esterase Enzyme

Sulfatase Enzyme

Neuraminidase Enzyme

Horseradish Peroxidase Enzyme

Stabilized 1,2-Dioxetanes

Our new series of ultra sensitive 1,2-dioxetanes can detect alkaline phosphatase enzyme at the Attogram level over a period of a few minutes.

Our Chemiluminescent system is at least 10 to 20 times more sensitive compared to the best competitor.

These novel 1,2-dioxetanes are protected by US Patent # 6,461,876 B1, 6,767,716 B2 and other pending PCT and US patent applications

Structure of the Molecule

Substrate for Alkaline Phosphatase

Advantages of Chemiluminescent Substrates for alkaline phosphatase

Formulated for two wavelength I max at 450nm and 540nm Safe – contains no organic solvents and no radioactive materials Unsurpassed signal – up to 15 times brighter than leading commercial reagents Low background – high signal to noise ratio allows increased signal without sacrifice Steady glow – long lasting light emission without signal decay for up to four hours Convenient – ready to use single bottle reagent requires no additional enhancers Consistent results – reproducible results and ideal for automatic or manual systems Very sensitive – less than one Attogram of alkaline phosphatase has been detected Wide dynamic range – 5 to 6 log dynamic range standard curves over a broad range of times Detection platforms – solution phase applications, membrane based applications and single tube or microplate format applications

The new series of stabilized chemiluminescent 1, 2-dioxetanes substrates can detect 10-23 moles of alkaline phosphatase in 100-microliter samples. These substrates are stable for two years at 4oC and can be stored at -20oC for an extended period of time.

>Single molecule detection was tested in microliter plate using Labsystem Fluoroskan Ascent FL luminometer. The enzyme used in this experiment was obtained from Sigma Chemical Company.

The synthesis of new 1, 2-dioxetanes can be shown as:-

Deuterium Based 1, 2-Dioxetanes

FIG. 1 - is a graph comparing the effect of isotopic hydrogens on the light emissions of a present 1, -2dioxetane (3) when compared to the 1,2-dioxetane of formula (10), when triggered by alkaline phosphatase enzyme in Tris buffer.

FIG. 1 - (Below) is a graph comparing the effect of isotopic hydrogens on the light emission of a present 1, 2-dioxetane (12) when compared to the 1, 2-dioxetane of formula (5), when triggered by alkaline phosphatase enzyme in Tris buffer.

FIG. 3 - is a graph comparing the effect of isotopic hydrogends on the light emmission of a present 1, 2-dioxetane (13) to [(4-Methoxy-4-(3-phosphoryloxy-4-chlorophenyl)] spiro [1,2-dioxetane-3 2'-adamantane], disodium salt (6), when triggered by alkaline phosphatase enzyme in Tris buffer.

FIG. 2 - is a graph comparing the effect of isotopic hydrogens on the light emission of a present 1, 2-dioxetane (15) to [4-Methoxy-4-(3-phosphoryloxy-4-chlorophenyl)] spiro [1, 2-dioxetane-3,2'-admantane], disodium salt (6), when triggered by alkaline phosphatase enzyme in Tris buffer.

FIG. 4 - is a graph comparing the effect of isotopic hydrogens on the light emission of a present 1, 2-dioxetane (16) to [4-Methoxy-4-(3-phosphoryloxy-5-methoxyphenyl)] spiro [1,2-dioxetane-3,2'-admantane], disodium salt (7), when triggered by alkaline phosphate enzyme in Tris buffer.

Luminol Based Stabilized Formulations for the Detection of Horseradish Peroxidase Enzyme

The new horseradish peroxidase substrates can detect horseradish peroxidase enzyme nearly to femtogram level in 100-microliter samples. These substrates are stable for eighteen months at 4oC as components A and B and can also be.