We added 100 µL of HUVEC suspension containing 1.0 × 104 cells into each well of a 96-well plate along with F-12K medium containing 100 µg/mL heparin and 2% fetal bovine serum and incubated the plate at 37°C for 5 hours. Next, 100 ng/mL VEGFA was neutralized by incubation with either IDB0062 or ranibizumab (3 hours, 37°C) before cell treatment. The VEGFA–IDB0062/ranibizumab mixture was added into each well, and the plate was incubated (72 hours, 37°C). Cell proliferation was measured using the WST-1 assay kit (DoGenBio, Seoul, Korea), according to the manufacturer's protocol.
Wst 1 assay kit
Manufactured by DoGenBio
The WST-1 assay kit is a colorimetric method for the quantitative determination of cell viability and proliferation. The assay relies on the cleavage of the tetrazolium salt WST-1 to formazan by cellular mitochondrial dehydrogenases. The amount of formazan dye generated is directly proportional to the number of metabolically active cells in the culture.
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Lab products found in correlation
2 protocols using wst 1 assay kit
1
HUVEC Proliferation Assay with VEGF Neutralization
2
Assessing Rat Glioma Cell Viability
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Cell viability was determined using water‐soluble tetrazolium salt‐1 (WST‐1) assay kit (DoGenBio). Rat C6 glioma cells were seeded into 96‐well plates (SPL Life Sciences) at density of 3 × 105 cells/well in 100 ml of media. Cells were incubated at 37°C in a 5% CO2 incubator overnight and then treated by PI extract at a final concentration of 0.06 μg/ml in triplicates for 24 hr. After that, 10 ml of EZ cytox cell viability assay reagent was added to each well followed by incubation at 37°C for 2 hr in a 5% CO2 incubator. Finally, the optical density was measured at 450 nm using a microplate reader (Sunrise™; Tecan). All experiments were repeated at least three times. Rat C6 glioma cells were also incubated with PI extract at different concentrations (6, 0.6 and 0.06 μg/ml), and the concentration that produced 50% decrease in cell viability (inhibitory concentration [IC50]) was calculated using GraphPad PRISM version 7.0.1 (GraphPad Software).
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