Principle of Assay
Cell movement and location studies require specialized probes that are nontoxic to living cells. This kit provides a versatile and well-retained cell tracing reagent (Green probe) for monitoring cell movement, location, proliferation, migration, chemotaxis, and invasion. The green probe can passively diffuse into cells and remain colorless and nonfluorescent until its acetate groups are cleaved by intracellular esterases to yield highly fluorescent, amine-reactive carboxyfluorescein succinimidyl ester. The succinimidyl ester group reacts with intracellular amines, forming fluorescent conjugates (Ex/Em= 494/521nm) that are well-retained and can be fixed with aldehyde fixatives. Excess unconjugated reagent and by-products passively diffuse to the extracellular medium, where they can be washed away. After conversion to permeant versions, the green probe is well retained in living cells through several generations and can display fluorescence for at least a week. The probe is transferred to daughter cells, but are not transferred to adjacent cells in a population.
