Two Parameter Cytotoxicity Test Kits

IN CYTOTOX products are available worldwide

NR test principle

The neutral red (NR) assay procedure is a cell survival/viability assay, based on the ability of viable cells to incorporate and bind neutral red within lysosomes. It is generally performed on adherent cells. NR is a weak cationic dye that readily penetrates the cell membrane and accumulates intracellularly in lysosomes (lysosomal pH < cytoplasmic pH), where it binds with anionic sites to the lysosomal matrix (G. GRIFFON, et al. 1995). Changes of the cell surface or the sensitive lysosomal membrane lead to lysosomal fragility and other changes that gradually become irreversible. Such alterations brought about by the action of xenobiotics result in a decreased uptake and binding of NR. It is thus possible to distinguish between viable, damaged, or dead cells, which is the basis of the assay.

The quantity of dye incorporated into cells is measured by spectrometry at 540 nm, and is directly proportional to the number of cells with an intact membrane.

The assay can be used to evaluate cytotoxicity by determination of the IC50 (50% inhibiting concentration).

CVDE test principle

This is a simple assay useful for obtaining quantitative information about the relative density of adherent cells in 96-well plates. After incubation with the test compound, the culture medium is discarded and the cells are washed to remove residual proteins of the culture medium which would interfere with the dye CVDE. CVDE is a dye that binds electrostatically to nuclear proteins and stains DNA.

After elimination of excess dye and solubilization of the fixed dye CVDE, the amount taken up by the cells can be measured spectrophotometrically at 540 nm and can be correlated to cell number.

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