Colorimetric cytotoxicity assay kit

Manufactured by Roche

Sourced in Italy, United States

The Colorimetric cytotoxicity assay kit is a laboratory equipment product that can be used to measure cell viability and cytotoxicity. It utilizes a colorimetric method to quantify the metabolic activity of cells.

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Lab products found in correlation

Tunel assay, by Roche (1 mentions) Crystal violet, by Merck Group (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)

Spelling variants of this product

Colorimetric assay (27 citations) Colorimetric assay kits (9 citations) Cytotoxicity assay kit (4 citations)

3 protocols using colorimetric cytotoxicity assay kit

Evaluating Cell Injury and Death

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Cell injury was assessed by measuring lactate dehydrogenase (LDH) release into the supernatants using a colorimetric cytotoxicity assay kit according to the manufacturer’s directions (Roche, Indianapolis, Indiana). The percentage LDH release was normalized to a no-stress vehicle control group for each cell line.
Cell death was evaluated using a TUNEL assay (Roche) per the manufacturer’s instruction. Cells were counterstained with Hoechst. Negative controls were set up per the manufacturer’s instructions, and DNAse-treated iCMs were used as a positive control. Coverslips were mounted, and cells were analyzed by fluorescent microscopy. A series of 10 images were taken from randomly selected areas on the coverslip. The TUNEL (+) nuclei in each image were quantified and compared against Hoechst-stained nuclei to give a percentage of TUNEL (+) cells.

Afzal M.Z., Reiter M., Gastonguay C., McGivern J.V., Guan X., Ge Z.D., Mack D.L., Childers M.K., Ebert A.D, & Strande J.L. (2016). Nicorandil, a Nitric Oxide Donor and ATP-Sensitive Potassium Channel Opener, Protects Against Dystrophin-Deficient Cardiomyopathy. Journal of cardiovascular pharmacology and therapeutics, 21(6), 549-562.

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Cellular Metabolic Activity Assay

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The ability of cells to reduce 3-(4,5-dimethyl thiazol-2-y1)-2,5-diphenyl tetrazolium bromide (MTT) was assessed as an index of cellular metabolic activity and mitochondrial integrity, as previously described [25 (link)]. MTT (Sigma, Saint Louis, MO, USA) was added at a final concentration of 0.25 mg/mL during the last 4 h of incubation. The medium was then removed, and 100 µL DMSO was added to each well to dissolve the dark blue crystals. The plates were then read on a microplate reader, using a test wavelength of 570 nm and a reference wavelength of 630 nm. The cell number in each condition was estimated by crystal violet (CV, Sigma, Saint Louis, MO, USA) dye [55 (link)]. Besides, to evaluate the membrane integrity, the release into the culture supernatant of the cytosolic enzyme lactate dehydrogenase (LDH) was measured using a colorimetric cytotoxicity assay kit (Roche Diagnostics, Milan, Italy).

Bernardo A., Malara M., Bertuccini L., De Nuccio C., Visentin S, & Minghetti L. (2021). The Antihypertensive Drug Telmisartan Protects Oligodendrocytes from Cholesterol Accumulation and Promotes Differentiation by a PPAR-γ-Mediated Mechanism. International Journal of Molecular Sciences, 22(17), 9434.

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Measuring iPSC-CM Cytotoxicity via LDH Assay

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Lactate dehydrogenase (LDH) activity was used as an assay of iPSC-CM cytotoxicity. iPSC-CMs were plated at 1×10⁴ cells/well in 96-well plates and cultured in 100 µL of RPMI/B27 medium supplemented with the appropriate glucose concentrations as previously discussed. LDH was measured via a colorimetric cytotoxicity assay kit according to the manufacturer’s directions (Roche Diagnostics Corporation, Indianapolis, IN, USA).

Canfield S.G., Zaja I., Godshaw B., Twaroski D., Bai X, & Bosnjak Z.J. (2016). High Glucose Attenuates Anesthetic Cardioprotection in Stem-Cell-Derived Cardiomyocytes: The Role of Reactive Oxygen Species and Mitochondrial Fission. Anesthesia and analgesia, 122(5), 1269-1279.

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