Streamlined Workflow for Cell Cycle Analysis

PerFix-nc Buffer R1, a formaldehyde-based reagent, is used to mildly fix the cells at room temperature followed by two consecutive washes allowing the removal of formaldehyde excess.

Permeabilization and co-staining of DNA and proteins is done simultaneously by addition of conjugate antibodies into the cell kit reagent containing Propidium Iodide and RNAseA, which ensures the removal of signal from staining RNA molecules.

There are no washes required following staining and the specimen is ready for acquisition after a 15 minute incubation.  This streamlined protocol reduces the number of steps to 7 from 11 and reduces the processing time by 50%.  Eliminating the post staining washes also reduces cell loss that could occur during washing steps, which could be important for rare event detection.

Workflow Comparison for Advanced Cell Cycle Staining. Schematic representation of the classical workflow versus the streamlined workflow utilizing PerFix-nc fixation reagent and the Cell Cycle Detection Kit.