Jc 1 dye buffer

JC-1 is an ideal fluorescent probe used in the detection of mitochondrial membrane potential. Early apoptosis is characterized by mitochondrial membrane potential drop, and fluorescence from red (aggregates) to green (monomer) by JC-1 shift can be easily detected as an apoptosis index of early detection. PMVECs were resuspended and centrifuged to collect and then incubated with JC-1 working solution (Beyotime Biotechnology, China) for 20 min at 37°C in the dark. PMVECs were washed by JC-1 dye buffer (Beyotime Biotechnology) and centrifuged at 600g and 4°C for 4 min. Finally, cells were incubated in 0.5 mL cold JC-1 dye buffer (1X, Beyotime Biotechnology) and transferred to a tube on ice for flow cytometry analysis by flow cytometry (ACEA NovoCyte). Analysis was carried out by NovoExpress software (ACEA NovoCyte).

MMP was determined by flow cytometry (FC 500 MPL; Beckman Coulter Inc., Fullerton, CA, USA) using 5,5ʹ,6,6ʹ-tetrachloro-1,1ʹ,3, 3ʹ-tetraethylbenzimidazol-carbocyanine iodide (JC-1) staining. JC-1 (C2006-1, Beyotime Institute of Biotechnology) is a lipophilic cationic dye that can selectively enter into mitochondria; its color reversibly changes with mitochondrial membrane depolarization. In normal cells with a high MMP, JC-1 spontaneously forms complexes (known as J-aggregates) and exhibits intense red fluorescence. In unhealthy cells with a low MMP, JC-1 remains in its monomeric form and exhibits green fluorescence.^{14 (link)} HL-60 cells were harvested and incubated with JC-1 at 37°C for 20 minutes at 1 × 10⁶ cells/mL. The cells were washed with JC-1 dye buffer (C2006-3, Beyotime Institute of Biotechnology) and analyzed using the FlowJo flow cytometry analysis software (FlowJo LLC, Ashland, OR, USA) at 525 nm excitation and 590 nm emission.