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In Vitro Mammalian Cell Gene Mutation Test Kit
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Cat.No.
GTK-M005
Description
The In Vitro mammalian cell gene mutation test can be used to detect TK gene mutations induced by chemical substances in suitable cell lines including L5178Y mouse lymphoma cells. The cells used are selected based on growth ability in culture and stability of the spontaneous mutation frequency. Cells deficient in thymidine kinase (TK) due to the mutation TK+/- -→ TK-/- are resistant to the cytotoxic effects of the pyrimidine analogue trifluorothymidine (TFT). Thymidine kinase proficient cells are sensitive to TFT, which causes the inhibition of cellular metabolism and halts further cell division. Thus mutant cells are able to proliferate in the presence of TFT, whereas normal cells, which contain thymidine kinase, are not.
Cells in suspension or monolayer culture are exposed to the test substance, both with and without metabolic activation, for a suitable period of time and subcultured to determine cytotoxicity and to allow phenotypic expression before mutant selection. Cytotoxicity is usually determined by measuring the relative cloning efficiency (survival) or relative total growth of the cultures after the treatment period. The treated cultures are maintained in growth medium for a sufficient period of time, characteristic of each selected locus and cell type, to allow near-optimal phenotypic expression of induced mutations. Mutant frequency is determined by seeding known numbers of cells in medium containing the selective agent to detect mutant cells, and in medium without selective agent to determine the cloning efficiency (viability). After a suitable incubation time, colonies are counted. The mutant frequency is derived from the number of mutant colonies in selective medium and the number of colonies in non-selective medium.
Cells in suspension or monolayer culture are exposed to the test substance, both with and without metabolic activation, for a suitable period of time and subcultured to determine cytotoxicity and to allow phenotypic expression before mutant selection. Cytotoxicity is usually determined by measuring the relative cloning efficiency (survival) or relative total growth of the cultures after the treatment period. The treated cultures are maintained in growth medium for a sufficient period of time, characteristic of each selected locus and cell type, to allow near-optimal phenotypic expression of induced mutations. Mutant frequency is determined by seeding known numbers of cells in medium containing the selective agent to detect mutant cells, and in medium without selective agent to determine the cloning efficiency (viability). After a suitable incubation time, colonies are counted. The mutant frequency is derived from the number of mutant colonies in selective medium and the number of colonies in non-selective medium.
Applications
Genotoxicity evaluation of food, drugs, chemicals, cosmetics, health care products, pesticides, medical devices etc.
Components
L5178Y Cells TK+/-clone (3.7.2C)
THMG (100x)
THG (100x)
TFT (100x)
S9 Mixture
MTT
Positive Controls
THMG (100x)
THG (100x)
TFT (100x)
S9 Mixture
MTT
Positive Controls
Size
20 mL*24 test/Kit
Storage
Store at -70°C.
Shipping
Dry ice
Citation Guidance
If you use this products in your scientific publication, it should be cited in the publication as: Creative Bioarray cat no. If your paper has been published, please click here to submit the PubMed ID of your paper to get a coupon.
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