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APPLICATIONS |
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LOW LEVEL DETECTION OF: |
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Alkaline Phosphatase Enzyme |
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β-Galactosidase Enzyme |
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β-Glucuronidase Enzyme |
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β-Glucosidase Enzyme |
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Esterase Enzyme |
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Sulfatase Enzyme |
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Neuraminidase Enzyme |
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Horseradish Peroxidase Enzyme |
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Stabilized 1,2-Dioxetanes |
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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. |
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Our Chemiluminescent system is at least 10 to 20 times more sensitive compared to the best competitor. |
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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 |
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Structure of the Molecule |
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Substrate for Alkaline Phosphatase |
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Advantages of Chemiluminescent Substrates for alkaline phosphatase |
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- 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 sacrafice
- Steady glow - long lasting light emission with out signal decayfor 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 platfroms - solution phase applications, membrane based applications and single tube or microplate format applications
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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. |
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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. |
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The synthesis of new 1, 2-dioxetanes can be shown as: |
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Deuterium Based 1, 2-Dioxetanes |
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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. |
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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. |
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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. |
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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. |
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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. |
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Luminol Based Stabilized Formulations for the Detection of Horseradish Peroxidase Enzyme |
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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. |