Neutrophils of healthy donors were stimulated by PMA (25 nM) for 2.5 h, followed by the addition of the impermeable SYTOX Green DNA dye (Thermo Fisher, USA, Cat No. S7572) and the viable cell dye Hoechst 33342 (Beyotime, China, Cat No. C1025) at a volume ratio of 1 : 100 per well. NET production in each well was observed under an Olympus fluorescence microscope (Japan), and the randomly selected fields were photographed.
Sytox green dna dye
Manufactured by Thermo Fisher Scientific
Sourced in United States
SYTOX Green DNA dye is a nucleic acid stain used for detecting dead cells in a sample. It is a high-affinity, green-fluorescent dye that is membrane-impermeant, allowing it to selectively stain cells with compromised plasma membranes.
Automatically generated - may contain errors
4 protocols using sytox green dna dye
1
Neutrophil NET Formation Assay
2
Apoptosis Detection by Flow Cytometry
3
Neutrophil NETosis Induction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neutrophils were isolated from freshly collected whole blood of healthy adults using the EasySep Direct Human Neutrophil Isolation Kit (STEMCELL Technologies, Vancouver, Canada) with greater than 95% purity. Neutrophils were resuspended at a concentration of 1 × 106 cells/mL in plasma isolated from healthy donors or fluid from chronic SDH for one hour. After one hour, the neutrophils were washed and resuspended in M199 at 37 °C. After 2 h, cell-free media was isolated, and NETosis was measured using SYTOX Green DNA dye (ThermoFisher), a dye that measures cell-free DNA.
4
Neutrophil Nuclear Kinetics by Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Neutrophils were resuspended at 2 × 106 cells/mL and incubated with NUCLEAR-ID® red DNA stain (1 µL per 1.5 × 106 cell suspension, Enzo Life Sciences, Farmingdale, NY, USA) for 5 min in the dark. Cells were washed 3 times in RPMI medium and spun down at 2400 g for 5 min. Stained cells were then plated at 20,000 cells per 100 µL in a 96-well poly-l-lysine coated flat bottom plate and incubated for 20 min to settle. Cells were then stimulated or not with PMA or ionomycin at given concentrations in a mixture with 2 µM SYTOX® Green DNA dye (Thermo Fisher, Cat No. S7020, Waltham, MA, USA) and were placed in the Incucyte® ZOOM platform contained within an incubator at 37 °C and 5% CO2.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!