The cytotoxicity of several cells was determined by Cell Count Reagent SF using WST-8 as a chromogenic substrate (Nacalai Tesque, Kyoto, Japan) [19 (link)]. The cells were seeded in 12-well plates and cultured with PBS or L. plantarum extract for 24, 48 and 72 h. At the end of treatment, the media were replaced with fresh medium and added mixed solution, including WST-8 and 1-Methoxy PMS. After incubation at 37 °C for 1 h, the media supernatants were measured at 450 nm using a microplate reader (Bio-Rad, Hercules, CA, USA).
Cell count reagent
Manufactured by Nacalai Tesque
Sourced in Japan
The Cell Count Reagent is a solution used to count cells in a sample. It provides a reliable and consistent method for determining the number of cells present in a given volume.
Automatically generated - may contain errors
4 protocols using cell count reagent
1
Cell Cytotoxicity Analysis via WST-8
2
Cell Viability Assay with WST-1 Reagent
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell viability was determined using the WST--1 reagent (Cell Count Reagent; Nacalai-Tesque) according to the manufacturer’s protocol. Target cells (ARPE-19 or hCMEC/D3) were plated at 1.5 × 104 cells/well in 96-well microtiter plates. ARPE-19 cells were treated for 24 h with NF-κB inhibitor or cytokines, or hCMEMC/D3 cells were treated with diluted conditioned media for 24 h. Cell viability reagent (10 μL) was added to each well for 4 h, and then, the resulting color change was measured using a Tecan-2000 plate reader at 450 nm wavelength. Relative changes in absorbance were determined after subtracting the absorbance measured in cell-free wells. Cell viability was then calculated as percent changes relative to DMSO-treated or control media-treated control cells.
3
Ruthenium Red and Tranilast Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RAW264.7 cells were seeded at 3.5 × 103 per well in 96-well plates, and pre-incubated without RANKL for 24 h. Then, several concentrations of ruthenium red (Sigma-Aldrich) or tranilast (Cayman Chemical) was added to each well. After 48 h of incubation, 10 μL of Cell Count Reagent (Nakarai Tesque, Kyoto, Japan) were added to each well. After reaction for 1 h, 10 μL of 0.1 M HCl was added to each well to stop the reaction, and the absorbance at 450 nm was measured using a microplate reader.
4
Cytotoxicity of CLE on RAW 264.7 and Macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytotoxicity of CLE to RAW 264.7 cells and mouse peritoneal macrophages was examined using a Cell Count Reagent (Nacalai Tesque) based on WST-8 according to the manufacturer’s instructions. Peritoneal macrophages and RAW 264.7 cells were seeded into a 96-well culture plate at 5.0 × 103 cells/well and precultured at 37 °C for 2 and 16 h, respectively, under humidified 5% CO2. After removing the medium, peritoneal macro-phages and RAW 264.7 cells were cultured in the medium containing various concentrations of CLE for 24 h. After removing the medium, the cells were stimulated with 1.0 µg/mL of LPS in the medium for 6 h at 37 °C. After washing with PBS, fresh medium containing 10% Cell Count Reagent was added to each well, and the absorbance was measured at 450 nm using an iMark microplate reader (Bio-Rad Laboratories, Hercules, CA, USA).
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!