Cell cytotoxicity / Proliferation assay cell type - HUVEC

Cell cytotoxicity assays measure the ability of certain compounds or chemical mediators to reduce the viability of the cells. The term cell cytotoxicity assay can sometimes be used interchangeably with cell proliferation assay. Healthy living cells can be identified by the use of formazan dyes, protease biomarkers or by measuring ATP content. The formazan dyes are chromogenic products formed by the reduction of tetrazolium salts by dehydrogenases, such as lactate dehydrogenase (LDH) and reductases that are released during cell death. Common tetrazolium salts include INT, MTT, MTS and XTT. Cell cytotoxicity can also be measured by using the SRB and WST-1 assays. These assays can usually be used in a high-throughput fashion and can be quantitated by measuring absorbance, colorimetry or luminescence. All these assays require similar numbers of cell plating at the initiation, a time course of treatment with the cytotoxic agent and at least triplicates for each condition at every point of analysis. Cell shrinkage, plasma membrane blebbing, cell detachment, externalization of phosphatidylserine, nuclear condensation and ultimately DNA fragmentation are well-described features of apoptosis. The assays that rely on cell membrane integrity for their function, may not be able to quantify early apoptosis. Therefore, in order to distinguish early apoptotic vs. late apoptotic or necrotic cells, additional flow cytometry techniques can be used. A combination of Annexin V and PI (propidium iodide) can be used to distinguish early (Annexin V+/PI-) and late apoptotic (Annexin V+/PI+) cells. Sometimes, caspase assays are used in order to differentiate the stages of apoptosis.

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Found 5 matching solutions for this experiment

Upstream tips
- cells were fixed with 2% paraformaldehyde for 20 min and permeabilized with 1% Triton X-100 in PBS for 5 min.

Protocol tips
- Cells were labelled for 16h.

- Cells were denatured with 2 N HCl for 30 min at 37°C,

- Cells were blocked for 30 min by using 3% BSA in PBS.

A&A Biotechnology

Protocol tips
RNA from total mice lungs were isolated using Fenozol (A&A Biotechnology). The quantity of ribosomal RNA and DNA contamination was examined using electrophoresis in 1% denaturing formaldehyde gel.
Upstream tips
- 1 × 104 cells/ml cells were platted in a 96 well plate.

Cells were pretreated with different concentrations of resveratrol
Protocol tips
- Cells were treated with 0.5 mg/ml MTT
Protocol tips
- Cells were incubated for 2 h at 37 °C after addition of CCK-8 reagent
Upstream tips
- Cells were seeded at a density of 5000 cells/cm2.
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