An amplified chemiluminescent assay for the detection of alkaline phosphatase.
An amplification assay for the measurement of alkaline phosphatase has been combined with a luminescent end point using the luminol-peroxidase system to produce the first enzyme-amplified chemiluminescent assay based on the principle of prosthetogenesis. This assay is both quantitative and extremely sensitive. When the assay was used to detect alkaline phosphatase in solution, the detection limit was 0.4 amol in a 5-min assay. The interassay variance ranged from 4 to 20% and 7 to 19% across the dynamic range of the assay for a chemiluminescent assay and an enhanced chemiluminescent assay, respectively, employing two different preparations of luminol.
Fisher
M
M
London Biotechnology Limited, Department of Biochemistry and Molecular Biology, University College London, United Kingdom.
Harbron
S
S
Rabin
B R
BR
D000469:Alkaline Phosphatase / Q000032:analysis* / Q000378:metabolism; D003605:D-Amino-Acid Oxidase / Q000378:metabolism; D004789:Enzyme Activation; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives* / Q000378:metabolism; D006735:Horseradish Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry*; D011392:Proline / Q000378:metabolism; D013997:Time Factors
D005182:Flavin-Adenine Dinucleotide; C078555:flavin-adenine dinucleotide phosphate; D008165:Luminol; D011392:Proline; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D003605:D-Amino-Acid Oxidase; D000469:Alkaline Phosphatase