Cell permeabilization and stabilization reagent and method of use

Abstract

A cell permeabilization and stabilization reagent and method of use are disclosed. The reagent contains a N-acyl sarcosine or a salt thereof, a pH adjusting agent to adjust pH of the reagent in a range from about 4 to about 6; and an aqueous medium; the reagent having a low ionic strength defined by a conductivity of less than 9.0 mS / cm. The reagent further contains bovine serum albumin and glycerol. The reagent may further include an alkyl sulfate surfactant. Upon incubating the cells with the reagent, the reagent permeates the cellular membrane to allow penetration of an intracellular marker, causes intracellular protein aggregation within the cellular membrane, while preserves a cellular constituent for binding with a cellular marker for subsequent analysis by flow cytometry.

Description

technical field

[0001]

[001] The present invention relates to a cell permeabilization and stabilization reagent and method of use thereof for preparing a cell-containing sample for analysis of cellular components. Background technique

[0002]

[002] It is an urgent requirement to analyze the internal components of cells at the molecular level. Several probes and antibodies have recently appeared, which are generally available for routine research and indicate intracellular structures. These probes and antibodies, because of their macromolecular nature, cannot permeate cells through cell membranes by themselves. Cells therefore need to be treated in order to render the cell membrane permeable (permeabilization grade). This treatment causes a major modification of the outer lipid membrane and, depending on the method used, can lead to loss of cell morphology, or even to the death of the entire cell.

[0003]

[003] Standard permeabilization methods involve treating cells by ...