MLO-Y4, MDA-MB-231, or RAW264.7 cells were seeded in 48-well plates (day 0) and treated with DMSO or Yoda1 (day 1). Cell seeding density differed between different durations of studies to avoid over confluence at the endpoint. After 2, 24, or 48 h incubation, cells were stained and incubated with 20 μL NucBlue® Live (Ex: 360 nm, Em: 460 nm, all Invitrogen, Waltham, MA, USA) and 20 μL NucGreen® Dead (Ex: 504 nm, Em: 523 nm, Invitrogen) stain for 30 min, following by capturing three random images per well using the fluorescence microscope (Nikon, Minato City, Tokyo, Japan). The images were analyzed using ImageJ to determine the number of cells. NucBlue® Live stained all cells, while NucGreen® Dead stained only dead cells. Positive control (PC) was prepared by treating cells with 70% ethyl alcohol for 15 min prior to staining.
Nucgreen dead
Manufactured by Thermo Fisher Scientific
Sourced in United States
NucGreen® Dead is a fluorescent nucleic acid stain used to detect dead cells. It provides a simple and reliable method to identify and quantify cell death in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Nucblue live, by Thermo Fisher Scientific (7 mentions)
Readyprobes cell viability imaging kit, by Thermo Fisher Scientific (2 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Axio observer z1 microscope, by Zeiss (1 mentions)
Mitotempo, by Merck Group (1 mentions)
Mitosox red mitochondrial superoxide indicator for live cell imaging, by Thermo Fisher Scientific (1 mentions)
Elesclomol, by MedChemExpress (1 mentions)
7 protocols using nucgreen dead
1
Assessing Cell Viability and Cytotoxicity
2
Hypoxia-Reoxygenation Assay in AC16 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AC16 cells, a hybrid cell model derived from the fusion of adult ventricular myocytes with stable, proliferating SV40 fibroblasts [32 (link)], were cultured in DMEM with 10 % FBS and penicillin/streptomycin (Thermo, #15140122). Cells were incubated overnight at 37 °C in DMEM with and without AICAR (1.25 mM) before the hypoxia/reoxygenation (H/R) challenge. Immediately prior to challenge, media was replaced with either DMEM (normoxia group) or Esumi buffer containing 137 mM NaCl, 12 mM KCl, 0.5 mM MgCl2, 0.9 mM CaCl2, 20 mM HEPES, and 20 mM 2-deoxy-d -glucose at pH 6.2 (H/R group). Cells were challenged with either normoxia (20 % oxygen, 5 % CO2) or hypoxia (1 % oxygen, 5 % CO2) for 4 h, and then media was exchanged with normoxic DMEM. After 30 min of normoxia, cells were stained with NucBlue Live (Hoechst 33342, Invitrogen) and NucGreen Dead (Sytox green, Invitrogen) according to manufacturer recommendations and visualized on a Cell Insight CX7 HCX Platform (Thermo) at 20× to quantify the ratio of living to dead cells. These cells were pelleted, and flash frozen for protein quantification.
3
Cell Viability Assessment of βTC3 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
βTC3 cells were labelled with NucBlue® Live and NucGreen® Dead reagents (R37609, Invitrogen) following the manufacturer’s protocol, and cell viability was assessed by photographing random field at 40× magnification using the Axio Observer Z1 microscope (Zeiss). In order to quantify the percentage of dead cells over total cells, the number of NucBlue® Live and NucGreen® Dead positive cells was counted (AG and AM). Mean values and standard deviations were evaluated on the basis of three independent experiments.
4
Vibration Effects on Osteocyte-Endothelial/Cancer Cell Co-cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MLO-Y4 osteocytes were seeded at 1500K cells/mL into the osteocyte channel. HUVEC endothelial cells at 2000K cells/mL or MDA-MB-231 cancer cells at 4000K cells/mL were seeded into the lumen channel. Cells in the platform received vibration (60 Hz, 0.3 g, 1 h/day for 3 days) or remained static. Negative control (NC) was prepared by treating cells with 70% ethyl alcohol for 10 minutes prior to staining. Cells were stained with 10% of NucBlue® Live (Ex: 360 nm, Em: 460 nm, Invitrogen, USA) and 10% of NucGreen® Dead (Ex: 504 nm, Em: 523 nm, Invitrogen, USA) in media and then incubated in a 37°C incubator for 30 minutes. Four random images were captured per channel using a fluorescence microscope (Nikon, Japan). The images were quantified by ImageJ.
5
Elesclomol-Copper Complex Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Elesclomol was obtained from MedChemExpress (https://www.medchemexpress.com/ ) and made fresh daily at a concentration ranging from 1–10 mM in dimethyl sulfoxide (DMSO). The E:C complex was formed by mixing equimolar and equivolume additions of Elesclomol and copper chloride (dissolved in H2O). MitoSOX Red mitochondrial superoxide indicator for live-cell imaging was obtained from Molecular Probes (Eugene, OR, USA); the mitochondrial superoxide scavenger MitoTEMPO was obtained from Sigma Aldrich (St. Louis, MO, USA); the mitochondrial membrane potential probe TMRE (tetramethylrhodamine, ethyl ester) was obtained from Biotium (Fremont, CA, USA); and the live cell nuclear stain, NucBlue Live, and the cell viability stain, NucGreen Dead, were obtained from ThermoFisher Scientific (Waltham, MA, USA). All solutions were used according to the manufacturer’s specifications.
6
Staining and Flow Cytometry of TMVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TMVs were isolated as described above and resuspended in filtered PBS. TMVs were then stained with either NucBlue Live or NucGreen Dead reagents (Ready Probes Cell Viability Imaging Kit, Thermofisher) for 2 hours at room temperature with gentle inversion, according to manufacturer’s specifications. Stained TMVs were centrifuged, washed with additional filtered PBS, and analyzed by flow cytometry.
7
Staining and Flow Cytometry of TMVs
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!