Process for measuring apoptosis

Details Process for measuring apoptosis Process for measuring apoptosis

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01) G01N 33/50 (2006.01)

Patent

CA 2311954

The invention relates to a method for quickly and easily measuring the apoptosis. Mammal cells are co-transfected with a plasmid coded for a fluorescent protein, and with a plasmid carrying an interested gene. The apoptosis is measured after incubation and mild fixation in which the DNA content of the cells is determined by means of DNA-binding colorants, and the portion of the transfected cells is determined by means of flow-through cytometry. The method can be used in order to identify substances which modulate the proapoptotic or antiapoptotic effect of genes or gene products.

L'invention concerne un procédé permettant de mesurer rapidement et simplement l'apoptose. Des cellules de mammifère sont cotransfectées avec un plasmide qui code pour une protéine fluorescente et également avec un plasmide qui porte un gène d'intérêt. Après incubation et fixation douce, l'apoptose est mesurée par détermination de la teneur en ADN des cellules, au moyen d'un colorant fixant l'ADN, et de la proportion de cellules transfectées, par cytométrie en flux. Ce procédé peut être mis en oeuvre pour l'identification de substances qui modulent l'effet pro-apoptotique ou anti-apoptotique de gènes ou de produits géniques.

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