Biomaterial Database

A new microassay for the determination of alkaline phosphatase activity in early mouse concepti. 1994

A Ahumada, and L Izquierdo

Departamento de Biología, Universidad de Chile, Santiago.

A new enzymatic assay for the determination of alkaline phosphatase (APase) activity in preimplantation mouse concepti is described. This method allows estimation of APase activity of concepti extracts using the fluorogenic substrate, 3,6-fluorescein diphosphate (FDP). For measuring APase activity, 0.1% Nonidet P-40 was used to solubilize the enzyme. Control assays showed that this procedure does not modify the enzyme activity. According to the Km obtained for APase from mouse concepti (between 1-2 microM), the initial concentration of FDP was 20 microM, which is 10 fold the Km. The assay sensitivity allows continuous recording of the product generated and a reliable determination in less than 20 min. Results show that APase activity in mouse concepti may be detected from the 2-cell stage, increasing exponentially towards the blastocyst stage.

UI	MeSH Term	Description	Entries
D004622	Embryo, Mammalian	The entity of a developing mammal (MAMMALS), generally from the cleavage of a ZYGOTE to the end of embryonic differentiation of basic structures. For the human embryo, this represents the first two months of intrauterine development preceding the stages of the FETUS.	Embryonic Structures, Mammalian,Mammalian Embryo,Mammalian Embryo Structures,Mammalian Embryonic Structures,Embryo Structure, Mammalian,Embryo Structures, Mammalian,Embryonic Structure, Mammalian,Embryos, Mammalian,Mammalian Embryo Structure,Mammalian Embryonic Structure,Mammalian Embryos,Structure, Mammalian Embryo,Structure, Mammalian Embryonic,Structures, Mammalian Embryo,Structures, Mammalian Embryonic
D005260	Female		Females
D005470	Fluorometry	An analytical method for detecting and measuring FLUORESCENCE in compounds or targets such as cells, proteins, or nucleotides, or targets previously labeled with FLUORESCENCE AGENTS.	Fluorimetry,Fluorometric Analysis,Analysis, Fluorometric
D000469	Alkaline Phosphatase	An enzyme that catalyzes the conversion of an orthophosphoric monoester and water to an alcohol and orthophosphate. EC 3.1.3.1.
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D013997	Time Factors	Elements of limited time intervals, contributing to particular results or situations.	Time Series,Factor, Time,Time Factor
D051379	Mice	The common name for the genus Mus.	Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus