Direct cytotoxic activity of FWGE was assayed by incubating 5 x 104 cells/well (96-well plates) in 100 μl culture medium with the indicated concentrations of FWGE for up to 72 hours at 37°C, 5% CO2. Cell viability was assessed using an MTS-based assay (Promega) according to the manufacturer’s instructions and compared to untreated controls. IC50 values were calculated by fitting the dose-response data to a dose–inhibition curve using GraphPad Prism software. Cytotoxicity of heat-inactivated FWGE (80°C, 90 minutes), proteinase K-treated FWGE (100 μg/ml, 37°C, 1 hour) and the protein fraction FWGP were assayed in the same way. Three replicate wells per condition were used in 3 independent experiments.
Mts based assay
Manufactured by Promega
The MTS-based assay is a colorimetric method used to measure cell proliferation, viability, and cytotoxicity. The assay utilizes the tetrazolium compound MTS, which is bioreduced by cells into a colored formazan product that can be quantified using a spectrophotometer.
Automatically generated - may contain errors
3 protocols using mts based assay
1
Cytotoxic Activity of FWGE
2
Evaluating Cytotoxicity of HM1.24-ETA' in Myeloma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BMSC were obtained from MNC isolated by ficoll density centrifugation of patient-derived bone marrow aspirates and subsequently cultured in R10+. For co-culture experiments, stromal cells were trypsinized and transferred to 96-well plates (0.5 × 104 cells/well), and MM cells (INA-6, 2 × 104 per well) were added the following day. Cells were treated with HM1.24–ETA′ at the concentrations indicated. For blocking experiments, the parental HM1.24-IgG1 antibody or a control antibody were added in molar excess. After 3 days of culture, stromal cell viability was analyzed by a colorimetric MTS-based assay (Promega). DNA synthesis of INA-6 cells under co-culture conditions was measured by [3H]-thymidine uptake. In brief, cells were pulsed with [3H]-thymidine (TdR; 1 μCi/well; specific activity, 5.0 Ci/mmol; Hartmann Analytic, Braunschweig, Germany) for 6 h. Subsequently, DNA was transferred onto glassfiber filters and counted in a β-scintillation counter (Perkin Elmer, Rodgau, Germany).
3
Evaluating HUVEC Angiogenic Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The formation of vascular-like structures by HUVECs on growth factor-reduced Matrigel (BD Biosciences) was performed as previously described (20,27). Differentiation was quantified by measuring the area of the “tube-like” networks that form in three randomly chosen fields from each well. Each experiment was repeated three times. Chemotaxis of HUVECs was assessed by transwell assay with polycarbonate membranes coated with fibronectin (Corning)(3415) [25 (link)]. HUVECs were added to the upper chamber, and serum-deprived media supplemented with pemafibrate, FGF21 or vehicle was added to the lower chamber. Cells were allowed to migrate through the pores of the membrane for 10 hours. Cell proliferation was assessed by MTS-based assay (Promega)(G3580) [26 (link)]. HUVECs were stimulated with pemafibrate, FGF21 or vehicle for the indicated lengths of time under normoxic or hypoxic condition. Hypoxic conditions were generated using an AnaeroPack (5% O2, 5% CO2, Mitsubishi GAS Chemical)(3276LJ).
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!