Various assays are used for assessing cell proliferation, viability, and cytotoxicity. Examples include optical counting, [3H] thymidine, MTT, WST, lactate dehydrogenase (LDH) assay, and more. This selection guide aims to provide you with information that will allow you to pick the method most suitable for your specific experimental needs.
| Measure cell count and/or DNA content to reveal cell division and DNA replication, respectively. Methods: – Optical counting with hemocytometer – DNA quantification using fluorophores |
| Measure parameters of cellular health to determine the intracellular impact of cytotoxicity. Methods: – Measuring evidence of cellular metabolic activity such as esterase activity, ATP, NADH dehydrogenase activity via MTT or WST, and so on |
| Measure indicators of cell death to evaluate cytotoxicity. Methods: – Measure the activity of lactate dehydrogenase leaked out of damaged cell membranes – Use membrane-impermeable dyes to selectively stain inside cells with damaged membranes |
| 0:00 | Title |
| 1:11 | What are Cell Proliferation, Viability, and Cytotoxicity Tests? |
| 4:12 | Cell Viability: Comparison of Each Reagent and Assay Principle |
| 8:53 | Cytotoxicity: Comparison of Each Reagent and Assay Principle |
| 13:52 | Measurement of Multiple Parameters |
| 16:33 | More Detailed Cellular Function Analysis |
The MTT and WST techniques are widely-used methods for evaluating cell proliferation and cytotoxicity. They are valued for the simplicity of their respective procedures, their safety, and the reproducibility of their results. WST (water-soluble tetrazolium) was developed by DOJINDO Laboratories in the 1990’s and is now used worldwide in cell proliferation assays. Cell Counting Kit-8 (CCK-8) is a commercial product that utilizes the WST method with improved sensitivity. Cell Counting Kit-8 is the original WST product, with which more than 5,000 papers have been published worldwide.
| Product name | MTT | Cell Counting Kit-8 | ATP Assay Kit | Cell Counting Kit-F Calcein-AM |
|---|---|---|---|---|
| Target | Mitochondrial dehydrogenase activity | Cellular dehydrogenase activity | ATP | Esterase activity |
| Instrument | Microplate reader (colorimetric) | Microplate reader (colorimetric) | Microplate reader (luminometric) | Microplate reader, FCM, Fluorescence microscope (fluorometric) |
| Advantage | Low price | · Simple procedure · Highly stable reagent |
· High sensitivity · Short reaction time (15 min) · Simple procedure |
· Applicable for various instruments · Short reaction time (15 min) |
| Disadvantage | · Affected by reducing agents · Requires dissolution of reagent after reaction |
Affected by reducing agents | · Expensive · Half-life after light emission needs to be accounted for |
· Serum increases background · Leakage of reagent |
| Product code | M009 | CK04 | A550 | CK06 (kit) C396 (ready-to-use solution) C326 (powder) |
 Please watch this video for comparison between MTT and Cell Counting Kit-8. |
| Product name | Cytotoxicity LDH Assay Kit-WST | Cell Counting Kit-F (Calcein-AM) |
Propidium iodide (PI) | Trypan Blue |
|---|---|---|---|---|
| Target | Released Lacate dehydrogenase (LDH) | A marker (Calcein-AM) released into the supernatant (Calcein-Release Assay) | Cytoplasm(Live cell membranes are impermeable to PI, but not dead cell membranes) | Cytoplasm; cells taking up Trypan Blue are considered non-viable |
| Instrument | Microplate Reader (colorimetric) |
Microplate reader, FCM, Fluorescence microscope (Fluorometric) |
Fluorescence microscope, FCM | Microscope (colorimetric) |
| Advantage | ·Simple procedure ·Suitable for multi-sample assay |
·Convenient for cytotoxicity assay using 2+ kinds of cell samples | ·Low price ·Simple procedure |
·Low price ·No special instruments needed |
| Disadvantage | ·Increased background by serum | ·Leakage of reagent from living cells | ·Not applicable for multi-sample assay | ·False-positives due to cytotoxicity ·Not applicable for multi-sample assay |
| Product code | CK12 | CK06 (kit) C396 (ready-to-use solution) C326 (powder) |
P378 (ready-to-use solution) P346 (powder) |
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In order to verify the accuracy of results in a cytotoxicity assay, individual samples are measured according to multiple different principles. Cell Counting Kit-8 (code: CK04) and Cytotoxicity LDH Assay Kit-WST (code: CK12) measure dehydrogenase activity and the released amount of LDH as reflective of living and dead cells, respectively. As shown in the example below, separate measurements indicate that the number of living cells decrease, and the number of dead cells increase as a toxicant’s concentration increases.
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Cell Proliferation / Cell Cytotoxicity Assay Kits /Related Reagents
Cell Double Staning Kit /Live Cell Staining /Dead Cell Staining /Nuclear Staining /Mitochondria Staning /Tissue Staining /Nucleolus Staining /Lipid Droplet Staining /Cell Membrane Staining /Lysosome Staining
Reagents for Intracellular Calucuum Ion /Reagents for Intracellular Ion /Related Reagents
Protein Labeling Kits /Protein Labeling Reagents /HPLC Derivertization Reagents /Biotion Labeling Reagents /Related Reagents /Exsosome Labeling
Stress Maker Detection /NO Detection /NO Donor /NO Inhibitor /ACE Inhibition Assay /Reagents・Kits for Sulfur Biology /Antioxidant Assay Kit /Donors for Sulfur Biology
Bacterial Proliferation Assay Kit /Bacteria Staining /Bacterial Fluorescent Staining
Transfection Reagents /Nuclear Staining /Agarose /Related Reagents /Buffer for Molecular Biology
Detergents /Sets
Hetero-bifunctional Reagents /Homo-bifunctional Reagents /Others
Reductive Chromogenic Dyes /Electron Mediators /Oxidative Chromogenic Dyes /Trinder Reagents
Ionophores /Anion Eliminator /Solvent for Ion Electrode of Liquid Film Type
Buffers
EDTA /Other Chelator /Reagents for Chelator Titration
Chromogen/Metal Indicator
/Fluorine /Iron / /Water Hardness /Residual Chlorine /ABS /Cyan / /Chromium /Copper
AA Chelator /Related Reagents
Spectrozole /Luminazole / /Acnazole /Dehydration Solvent for Synthesis
Biochemicals
Alkanethiol Derivative /Phosphonic Acid Derivatives
