Enhancement of the sensitivity for in situ detection of alkaline phosphatase using a homopolymer of 2-acrylamide 2-methylpropanesulfonate.
In order to enhance the sensitivity of detection of alkaline phosphatase immobilized on nitrocellulose, a homopolymer of 2-acrylamide 2-methylpropanesulfonate (poly-AMPS) and agarose were added to the reaction mixture using 5-bromochloroindoxyl phosphate and nitroblue tetrazolium. By addition of the above ingredients, the amount of the reaction product deposited on the nitrocellulose filters on which alkaline phosphatase had been dot-blotted was increased, and darkening of the background (un-dot-blotted area) was prevented. Reflective densitometry revealed that the addition of poly-AMPS and agarose increased the sensitivity of the detection approximately three times higher than without the use of these compounds.
Yasue
H
H
Department of Animal Breeding and Genetics, National Institute of Animal Industry, Ibaraki, Japan.
Awata
T
T
D000178:Acrylamides / Q000494:pharmacology*; D000469:Alkaline Phosphatase / Q000032:analysis*; D000476:Alkanesulfonates / Q000494:pharmacology*; D003101:Collodion; D004800:Enzymes, Immobilized; D007211:Indoles / Q000494:pharmacology; D009580:Nitroblue Tetrazolium; D011108:Polymers
D000178:Acrylamides; D000476:Alkanesulfonates; D004800:Enzymes, Immobilized; D007211:Indoles; D011108:Polymers; D009580:Nitroblue Tetrazolium; C035455:5-bromo-4-chloro-3-indoxyl phosphate; C056288:2-acrylamido-2-methylpropanesulfonate; D003101:Collodion; D000469:Alkaline Phosphatase