Assay Time: 30min
Test Principle: Elabscience? Calcein AM/PI Double Staining Kit can be used to distinguish dead cells and living cells in mammals with esterase activity. Calcein AM is the addition of acetyl methoxy methyl ester (AM) group to traditional Calcein, which increases hydrophobicity and can easily penetrate the living cell membrane and enter the cell. Calcein AM itself has no fluorescence. After entering the cell, it is hydrolyzed by endogenous esterase in the cell to produce Calcein, a polar molecule with strong negative charge and cannot be retained in the cell through the cell membrane, while Calcein can emit strong green fluorescence ( Ex / Em = 494nm / 517nm ). Due to the lack of esterase, dead cells cannot or rarely produce Calcein, so only living cells are stained with strong green fluorescence, and dead cells cannot be stained or stained very weakly. The selective membrane permeability of dead cells is lost, and Propidium Iodide ( PI ) can enter the cell to specifically bind to double-stranded DNA and produce strong red fluorescence(Ex/Em = 535nm/617nm) to label dead cells. Therefore, the combination of Calcein AM and PI can perform double fluorescence staining on living cells and dead cells at the same time, which can be used for the detection of cell activity and cytotoxicity.
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