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Medicilon’s Cell Viability Assay

2016-12-12
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Medicilon Cell Viability Assay offers a simple, rapid, reliable, sensitive, safe and cost-effective measurement of cell viability.

Cell viability is a determination of living or dead cells, based on a total cell population. Cell viability assess healthy cells in a sample, with no distinction between dividing or quiescent cells. An increase in cell viability indicates cell growth, while a decrease in viability can generally be interpreted as the result of either toxic effects of compounds/agents or sub-optimal culture conditions.

Medicilon’s Cell Viability Assay

The goal: To distinguish live cells from dead and apoptotic cells in order to calculate the percentage of viable cells for each experiment.

Cell Viability Assay Kits
Cell Viability Assay Kits are a family of fluorescence-based reagents for the assessment of cell viability, cell proliferation and various apoptosis events using mammalian cells. Optimized for use with microplate readers, these assay kits employ a no-wash, homogeneous assay protocol that enables characterization of a full concentration-response profile of test compounds.

MTT Cell Viability Assay
MTT Cell Viability Assay Kit provides a highly sensitive, rapid and easy-to-use tool for detection of cytotoxic agents present in cell cultures. Determination of live cell numbers is often used to assess the rate of cell proliferation and cytotoxicity caused by drugs and cytotoxic agents. Among all non-radioactive viability assays, MTT assay developed by Mossman is one of the most versatile and popular assays. MTT is a tetrazolium salt that is turned into a purple formazan product after reduction by mitochondrial enzymes that are only present in metabolically active live cells, not in dead cells. The amount of formazan product generated is proportional to the number of living cells in the sample. The formazan product can be solubilized and then photometrically quantified at 570 nm.

Experimental Protocol
1. Plate cells into 96-well tissue culture plates. In general, cells should be seeded at densities between 5000 and 10,000 cells per well since they will reach optimal population densities within 48 to 72 hours.
2. Carry out your experiment by adding chemicals or biological agents into appropriate well. The final volume of tissue culture medium in each well should be 0.1mL, and the medium may contain up to 10% Fetal Bovine Serum.
3. Thaw one vial of MTT solution for each 96-well plate assay.
4. Add 10µL MTT solution to each well. Mix by tapping gently on the side of the tray or shake briefly on an orbital shaker.
5. Incubate at 37°C for 4 hours. At high cell densities (>100,000 cells per well) the incubation time can be shortened to 2 hours.
6. Add 200µL DMSO into each well to dissolve the formazan by pipetting up and down several times.
7. Measure the absorbance on an ELISA plate reader with a test wavelength of 570 nm and a reference wavelength of 630 nm to obtain sample signal.

Advantages:
Easy-to-use: There is no requirement for additional reagents and/or the cell washing procedures
Speed: Multiwell plates and an ELISA reader can be used for reading
Sensitivity: Can be assayed even in low cell concentrations
Accuracy: Dye absorbance is proportional to the number of cells in each well
Safety: There is no need for radioactive isotopes

MTS Cell Viability Assay
A Cell Viability Assay, often used to assay toxicity of compounds, that depends upon the production of formazan, a reduction product of MTS, by live cells. The intensity of the color (at 492 nm) due to formazan is proportional to the number of live cells. An alternative to the MTT assay in which formazan is produced from MTT by a mitochondrial dehydrogenase, which only happens in live cells.

Cell Viability Assessment

1) In addition to multi-parametric cytotoxicity assessment using high content screening, Cyprotex offer general cell viability assessment using single endpoints such as LDH (lactate dehydrogenase), neutral red or MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide).
2) LDH is a stable enzyme, present in all cell types, and is rapidly released into the cell culture medium upon damage of the plasma membrane. Neutral red measures the ability of viable cells to incorporate and bind the neutral red dye in the lysosomes. MTT is a yellow, water-soluble tetrazolium dye that is converted by mitochondrial dehydrogenases in viable cells to a water-insoluble, purple formazan.
3) LDH, neutral red and MTT are well established, sensitive and reliable endpoints of cytotoxicity.
4) Cyprotex offer assessment of LDH, neutral red or MTT in HepG2, NIH3T3, HaCaT cells or primary hepatocytes.

Contact Us 

Email : marketing@medicilon.com.cn
Tel : +86 021 58591500

Tips : Above is part of cell viability assay and cell viability test protocol. You can also CONTACT US with any question or enquiry you may have. We will be happy to discuss your needs in detail and design an appropriate plan of action.

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