Calcein-AM/PI，Live/Dead Cell Double Stain Kit

Product Introduction: Calcein-AM is a cell staining reagent for fluorescent labeling of living cells, emitting green fluorescence (Ex=490nm, Em=515nm). The addition of acetomethoxymethyl ester (AM) group to traditional Calcein increases its hydrophobicity and enables it to easily penetrate the membrane of living cells. Once inside the cell, calcein-AM (which is not fluorescent on its own) is cleaved by intracellular esterases to form the nonpermeable polar molecule Calcein, which is trapped inside the cell and emits strong green fluorescence. Compared with other similar reagents such as BCECF-AM and CFDA, AM is the most suitable fluorescent probe for live cell staining due to its very low cytotoxicity and does not inhibit any cellular functions such as proliferation and lymphocyte chemotaxis. Because dead cells lack esterase, Calcein-AM is used only for cell viability testing and short-term labeling of living cells. Therefore, Calcein-AM is often used in combination with dead cell fluorescent probes such as propidium iodide (PI) for simultaneous fluorescent double staining of live and dead cells. Propidium iodide (PI) can not pass through the cell membrane of living cells, but can only pass through the disordered region of the dead cell membrane to reach the nucleus and embed into the DNA double helix of the cell to produce red fluorescence (Ex=535 nm, Em=617 nm). Therefore, PI only stains dead cells. Since both Calcein and PI-DNA can be excited by 490 nm, both live and dead cells can be visualized by fluorescence microscopy. However, only dead cells could be observed with 545 nm excitation. The working principle of this kit is to double dye Calcein-AM and PI to double stain and label live and dead cells, so as to analyze the level of live and dead cells. According to our optimized experimental system, 200μl cell suspension can be stained for 500 times Note: Product information may be optimized and upgraded. Please refer to the actual label information for accuracy.