Viable cells that active mitochondria reductase which produce the purple formazan crystals\n
MTT AssayPurpose: To understand how theMTT asay works, what it measures, and why it is important.
How it works:The MTT assay is a colorimetric assay systemand is a quantitative method used to determinecell growth and viability.
Why is the MTT assay important: measuring cellular proliferation (cell growth) in response to external factors
Applications:•Drug sensitivity•Cytotoxicity•Response to growth factors•Cell activation
Yellow MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, atetrazole) is reduced to purpleformazan in the mitochondria ofliving cells. (Formazan dyes areartiﬁcial chromogenic substrates fordehydrogenases and reductases).
Actinomycin D is a substance that binds to DNAand prevents transcription.In this assay it serves as a negative control byinhibiting WEHI cell replication during theincubation time and ensures maximum assaysensitivity.
Count Adjust count cells to 2x105cells/ mL WEHI cells, Mɸ cells, use use RPMI- RPMI-1640 WEHI WEHI+ActD