Polymer toxicity was evaluated using human intestinal epithelial Caco-2 cells and the CytoTox-ONE assay kit (Promega), which measures the release of lactate dehydrogenase (LDH) from membrane-damaged cells, as previously described.27 (link) Briefly, approximately 1 × 104 cells in DMEM medium, containing 10% serum and other nutrients, were seeded in each well of a 96-well plate, which was incubated for 72 h at 37°C. Medium was exchanged for fresh DMEM (phenol red- and pyruvate-free) shortly before the assay was performed. Cells were treated with polymers at varied concentrations in a 2-fold serial dilution series ranging from 500 to 3.9 μg/mL for 6 h at 37°C. The cells in each well were then analyzed using the CytoTox-ONE assay kit. Caco-2 cells in DMEM only (without polymer) and cells treated with lysate solution to cause 100% release of LDH (10% Triton-X 100) were incorporated as the “blank” and positive control, respectively. Fluorescence intensity was measured on a microplate reader using ex/em 560/590 nm. Cell death was calculated using (% cell death = (Fpolymer − Fblank)/(Fpos.control − Fblank) × 100) and plotted against the polymer concentration. The IC10 value represents the polymer concentration needed to cause 10% cell death.
Cytotox one assay kit
Manufactured by Promega
Sourced in United States
The CytoTox-ONE assay kit is a fluorometric method for quantifying the release of lactate dehydrogenase (LDH) from damaged cells. It provides a convenient way to measure cytotoxicity or cell lysis in a variety of cell types.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using cytotox one assay kit
1
Quantifying Polymer Toxicity on Caco-2 Cells
2
Evaluating Polymer Toxicity on 3T3 Fibroblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Polymer toxicity was evaluated
using NIH 3T3 fibroblasts and the CytoTox-ONE assay kit (Promega),
which measures the release of lactate dehydrogenase (LDH) from membrane-damaged
cells, as described previously.34 (link) Briefly,
1.5 × 104 cells in DMEM were seeded in each well of
a 96-well plate, which was incubated for 24 h at 37 °C. Medium
was exchanged for fresh DMEM (phenol red- and pyruvate-free), and
cells were incubated for another 2 h at 37 °C. Cells were treated
with nylon-3 polymers at varied concentrations in a 2-fold serial
dilution series ranging from 400 to 3.13 μg/mL for 12 h at 37
°C. The cells in each well were then analyzed using the CytoTox-ONE
assay kit. On the same plate, wells without polymer and wells treated
with lysate solution to cause 100% release of LDH were incorporated
as the blank and positive control, respectively. Fluorescence intensity
was measured on a Tecan Infinite M1000 microplate reader using ex/em
560/590 nm. Cell death was calculated from (% death = (Fpolymer – Fblank)/(Fcontrol – Fblank) × 100) and plotted against polymer concentration. The IC10 value is the polymer concentration that causes 10% cell
death.
using NIH 3T3 fibroblasts and the CytoTox-ONE assay kit (Promega),
which measures the release of lactate dehydrogenase (LDH) from membrane-damaged
cells, as described previously.34 (link) Briefly,
1.5 × 104 cells in DMEM were seeded in each well of
a 96-well plate, which was incubated for 24 h at 37 °C. Medium
was exchanged for fresh DMEM (phenol red- and pyruvate-free), and
cells were incubated for another 2 h at 37 °C. Cells were treated
with nylon-3 polymers at varied concentrations in a 2-fold serial
dilution series ranging from 400 to 3.13 μg/mL for 12 h at 37
°C. The cells in each well were then analyzed using the CytoTox-ONE
assay kit. On the same plate, wells without polymer and wells treated
with lysate solution to cause 100% release of LDH were incorporated
as the blank and positive control, respectively. Fluorescence intensity
was measured on a Tecan Infinite M1000 microplate reader using ex/em
560/590 nm. Cell death was calculated from (% death = (Fpolymer – Fblank)/(Fcontrol – Fblank) × 100) and plotted against polymer concentration. The IC10 value is the polymer concentration that causes 10% cell
death.
3
Exosome and Oxidative Stress Impacts on NHDFs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NHDFs were cultured in the presence of exosomes and oxidative stress molecules, and the cells and culture medium were recovered at various times. The total number of cells and the number of surviving cells were counted using Countess cell counting chamber slides (Thermo Fisher Scienti c K. K. Tokyo, JAPAN) and a CytoTox-ONE Assay Kit (Promega, Madison, WI, USA), respectively (n = 3/group).
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!