ROS levels were evaluated using a ROS Assay Kit (R252; Dojindo, Kumamoto, Japan). The cells were plated in an 8-well 0.8 cm2 Lab-Tek chamber slide (177445PK; Thermo Fisher Scientific). After 24 h, the cells were washed with phenol red-free Hanks’ balanced salt solution (HBSS, 084-08965; Wako) and incubated in DCFH-DA solution and 0.5 μg/mL Hoechst 33342 (H342; Dojindo, Tokyo, Japan) for 30 min at 37 °C with 5% CO2. Thereafter, the cells were washed with HBSS and treated with H2O2 and IL/TNF for 30 min. After washing with HBSS, the fluorescence signal was detected using a fluorescence microscope (BZ-X700, KEYENCE).
Hanks balanced salt solution (hbss)
Manufactured by Keyence
Sourced in Japan
The HBSS (Hank's Balanced Salt Solution) is a laboratory reagent used for maintaining the physiological balance of cell cultures. It provides a balanced salt solution that supports the survival and growth of cells in vitro. The HBSS formulation includes essential inorganic salts, glucose, and other components to maintain the osmotic pressure and pH of the cell culture environment.
Automatically generated - may contain errors
Lab products found in correlation
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (3 mentions)
Hanks balanced salt solution (hbss), by Fujifilm (2 mentions)
Ferroorange, by Dojindo Laboratories (2 mentions)
Lipiradical green, by Funakoshi (2 mentions)
Ferroorange, by Keyence (2 mentions)
Lipiradical green, by Keyence (2 mentions)
Bz x800, by Keyence (2 mentions)
Lab tek chamber slide, by Thermo Fisher Scientific (1 mentions)
Ros assay kit, by Dojindo Laboratories (1 mentions)
Hoechst 33342, by Dojindo Laboratories (1 mentions)
Bz x700, by Keyence (1 mentions)
Mitosox, by Thermo Fisher Scientific (1 mentions)
Bz 8000 fluorescence microscope, by Keyence (1 mentions)
Glass bottom dishes, by Matsunami (1 mentions)
Liperfluo, by Dojindo Laboratories (1 mentions)
Bz 2 viewer software, by Keyence (1 mentions)
5 protocols using hanks balanced salt solution (hbss)
1
ROS Measurement in Cultured Cells
2
Intracellular Fe2+ and Lipid Radicals Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Detection of intracellular Fe2+ was performed using FerroOrange (excitation: 561 nm, emission: 570–620 nm) (Dojindo). Cells were cultured in glass-bottomed dishes for 3 d and washed with HBSS (Thermo Fisher Scientific), after which 1 μM FerroOrange was added for 30 min at 37°C, followed by observation under a microscope (BZ-X800; Keyence). Lipid radicals were detected by LipiRADICAL Green (Funakoshi) (excitation: 470 nm, emission: 520–600 nm). Cells were cultured in glass-bottomed dishes for 3 d and washed with HBSS, after which 1 μM LipiRADICAL Green was added for 10 min at 37°C, followed by observation under a microscope (BZ-X800; Keyence).
3
Intracellular ROS Visualization in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular •OH were detected using HPF (hydroxyphenyl fluorescein; Goryo Chemical Inc., Sapporo, Japan) and mtROS were detected using MitoSOXTM (Thermo Fisher Scientific) as recommended by the manufacturer. Briefly, HeLa and SAS CRR cells were cultured overnight in glass bottom dishes (Matsunami Glass Ind., Ltd., Osaka, Japan). Then, the cells were washed twice using Hank’s balanced salt solution (HBSS) (Fujifilm Wako) to remove residual FBS. For •OH, cells were treated with 10 µM of HPF in HBSS for 15 min at 37 °C. For mtROS, cells were treated with 5 µM of MitoSOXTM in HBSS for 10 min at 37 °C. After these treatments, cells were washed with HBSS three times and fluorescence images were obtained using a BZ-8000 fluorescence microscope (KEYENCE Corporation, Osaka, Japan) from 3 separate dishes for each treatment.
4
Intracellular Iron and Lipid Radical Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Detection of intracellular Fe2+ was performed using FerroOrange (excitation, 561 nm; emission, 570–620 nm) (DOJINDO, Kumamoto, Japan). Cells were cultured in glass-bottomed dishes for 3 days and washed with HBSS (Thermo Fisher), after which 1 µM FerroOrange was added for 30 min at 37°C, followed by observation under a microscope (BZ-X800; KEYENCE). To detect Fe2+ in mitochondria, 5 µM Mito-FerroGreen working solution (excitation, 488 nm; emission, 500–550 nm) (DOJINDO, Kumamoto, Japan) was added to the cells for 30 min at 37°C, followed by observation under a microscope. Lipid radicals were detected using LipiRADICAL Green (Funakoshi, Tokyo, Japan) (excitation, 470 nm; emission, 520–600 nm) and Liperfluo (excitation, 524 nm; emission, 535 nm) (DOJINDO, Kumamoto, Japan). Cells and cardiomyocyte were cultured in glass-bottomed dishes for 3 days and washed with HBSS, after which 1 µM LipiRADICAL Green or 4 µM Liperfluo working solution was added for 10 min at 37°C, followed by observation under a microscope (BZ-X800; KEYENCE).
5
Neutrophil Chemotaxis Assay
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!