Recycling cultured cells for immunofluorescent labeling

A method to use sequential rounds of immunofluorescent labeling in cell cul tures is presented. The method is based on the utilization of a non-liquid reducing agent, sodium dithionite, in conjunction with ionic or non-ionic d etergents (SDS or TX100, respectively) at room temperature. This method pre serves cell morphology and substrate antigenicity, and operates through the complete extraction of most primary and secondary antibodies. Using this p rotocol, the sequential immunolocalization of different proteins is possibl e, without signal interference with previous immunolabeling rounds. In addi tion, the method is also useful to recycle blotted membranes in immunoblots .