Enzymatic assay for the quantitative determination of...

Details Enzymatic assay for the quantitative determination of... Enzymatic assay for the quantitative determination of...

C - Chemistry, Metallurgy

12

Q

C12Q 1/46 (2006.01) C12Q 1/44 (2006.01) G01N 33/543 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2751962

The invention relates to a new method for measuring a calcium-dependent phospholipase A1 or A2 enzymatic activity in a sample, comprising the following steps: a) contacting said sample with a solid phase coated with a fluorochrome-labelled phospholipase A1 or A2 substrate, wherein the molecular coverage is in the range 8 to 30 fluorochrome-labelled phospholipase A1 or A2 substrate molecules / nm2 in conditions where calcium-dependent phospholipase A1 or A2 enzymatic activity is blocked. b) reading the fluorescence emission overtime c) adding a solution to initiate calcium-dependent phospholipase A1 or A2 enzymatic activity d) reading the fluorescence emission overtime, and kits for carrying out said method.

L'invention concerne un nouveau procédé de mesure d'une activité enzymatique phospholipase A1 ou A2 dépendante du calcium dans un échantillon, qui comprend les étapes suivantes : a) la mise en contact dudit échantillon avec une phase solide revêtue avec un substrat phospholipase A1 ou A2 marqué par un fluorochrome, la couverture moléculaire étant dans la plage allant de 8 à 30 molécules de substrat phospholipase A1 ou A2 marqué par un fluorochrome/nm2, dans des conditions dans lesquelles l'activité enzymatique phospholipase A1 ou A2 dépendante du calcium est bloquée ; b) la lecture de la prolongation de l'émission de fluorescence ; c) l'ajout d'une solution pour initier l'activité enzymatique phospholipase A1 ou A2 dépendante du calcium ; d) la lecture de la prolongation de l'émission de fluorescence. L'invention concerne également des kits pour réaliser ledit procédé.

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