multiparametric analysis of complex cellular response

To introduce a protocol for a simultaneous flow cytometric detection of two CD surface antigens, DNA synthesis and quantification, histone H2AX phosphorylation (marker for DNA damage), cleaved PARP (marker for apoptosis) and viability on representative panel of cell lines derived from human and mouse.
Conclusion:
We introduced protocol for a simultaneous flow cytometric detection of two CD surface antigens, DNA synthesis and quantification, histone H2AX phosphorylation (marker for DNA damage), cleaved PARP (marker for apoptosis) and viability on representative panel of cell lines derived from human and mouse. This method is useful tool for complex analysis of cells. For compensation setup were used anti-rat and anti-hamster Ig kappa/negative control compensation beads (BD Biosciences), SpheroTM Biotin Polystyrene Particles (Spherotech, Lake Forest, IL, USA) labeled with antibodies (Table S2) in appropriate dilution. ArcTM Amine Reactive Compensation Bead Kit beads (Thermo Fisher Scientific) were stained with LIVE/DEAD® Fixable fluorescent reactive dye kit. As compensation controls for DNA stain, fixed and permeabilized cells with/without appropriately diluted DNA probe were used. Compensations were calculated using FACSDiva software (Version 6.1.3; BD Biosciences)