CyQUANT™ MTT and XTT Cell Viability Assays
CyQUANT™ MTT and XTT Cell Viability Assays
Invitrogen™

CyQUANT™ MTT and XTT Cell Viability Assays

CyQUANT MTT and XTT Cell Viability assays are colorimetric microplate assays for accurate assessment of cell health, proliferation, and cytotoxicity. CyQUANT MTT is a well-established assay, while CyQUANT XTT offers improved performance and continuous measurement without cell lysis or solubilization.
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Número de catálogoTipo de coloranteEmission
V13154Otras etiquetas o colorantes
X12223XTT
Número de catálogo V13154
Precio (EUR)
481,00
1,000 assays
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Tipo de colorante:
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Precio (EUR)
481,00
1,000 assays
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CyQUANT MTT and XTT Cell Viability assays are colorimetric microplate assays used to measure the metabolic activity of cells, which is an indicator of cell viability, proliferation, and cytotoxicity. CyQUANT MTT and XTT function as cell health indicators by using the cellular redox potential of live cells to convert a tetrazolium-based salt reagent, MTT or XTT, to a brightly colored formazan product. The absorbance of the formazan can be measured using a microplate reader and correlates with the number of metabolically active cells to quantitively measure cell viability and proliferation.

Measuring cell viability is essential for assessing cell health, determining genotoxicity, and evaluating anti-cancer drugs. Colorimetric indicators such as MTT and XTT offer rapid, cost-effective, and consistent methods for quantitatively determining mammalian cell viability using microplate readers. The CyQUANT MTT assay is a well-established and popular cell viability assay, while the CyQUANT XTT assay provides a high dynamic range and low variability. In addition to improved performance, the CyQUANT XTT assay offers continuous measurement without the need for cell lysis or solubilization.

Features of the CyQUANT MTT Cell Viability Assay

  • Complete and optimized MTT assay kit—includes all reagents needed for colorimetric detection of viable mammalian cells
  • Trusted MTT reagent—uses the well-established MTT reagent to measure cellular redox potential
  • Configured to perform 1,000 tests using a 96-well microplate

Principle of the CyQUANT MTT Cell Viability Assay

The CyQUANT MTT assay is based on the reduction of the water-soluble, yellow tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) to insoluble, purple formazan crystals by metabolically active cells. The redox potential in the mitochondria of active mammalian cells reduces MTT to a strongly pigmented formazan product. After solubilization, the absorbance of the formazan can be measured on a microplate reader, which correlates with the number of metabolically active cells.

Protocol for the CyQUANT MTT Cell Viability Assay

Prepare the MTT reagent and incubate cells for a specific period, during which viable cells convert MTT into insoluble purple formazan product. The formazan crystals are then solubilized with the included SDS (sodium dodecyl sulfate) reagent or DMSO, and the absorbance is measured using a microplate reader at 570 nm.

Features of the CyQUANT XTT Cell Viability Assay

  • High reproducibility—absorbance-based viability assay that measures redox potential in live cells with low variability
  • Improved dynamic range and sensitivity—enhanced performance compared to other colorimetric viability assays
  • Easy mix-and-read format—configured for one 96-well plate at a time
  • Non-toxic and continuous—allows multiple time points to be collected without cell lysis
  • Quick and simple—no solubilization step required, making it faster and easier to perform than the MTT assay
  • Complete XTT assay kit—includes all the materials needed for ten 96-well plates

Principle of the CyQUANT XTT Cell Viability Assay

The CyQUANT XTT assay relies on the reduction of the water-soluble, yellow tetrazolium salt XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) to a water-soluble orange formazan product by metabolically active cells. Unlike MTT, XTT does not form insoluble crystals and produces a soluble formazan product that does not require solubilization. Since no cell lysis or solubilization are required for detection, the XTT assay can be used as a continuous assay, and multiple time points can be analyzed using an absorbance-based microplate reader.

In addition to the XTT Reagent, the CyQUANT XTT Cell Viability Assay kit includes electron coupling reagent to improve the dynamic range of the assay. The sensitivity and consistency of the assay is significantly increased when the electron coupling reagent is used with the CyQUANT XTT assay kit.

Protocol for the CyQUANT XTT Cell Viability Assay

For each 96-well plate, one bottle of XTT reagent and one tube of electron coupling reagent are thawed and mixed to create a stock solution, then added to cells. Cells are incubated for 2–4 hours, and metabolically active cells reduce XTT to form a water-soluble orange formazan dye. The absorbance of the dye is then measured using a microplate reader at 450 nm and 650 nm.

The XTT/electron coupling reagent stock solution should be used promptly after mixing, as extended room temperature storage or freeze/thaw cycles reduce assay performance and sensitivity. To ensure optimal performance and reproducibility, the CyQUANT XTT assay kit consists of separate XTT and electron coupling reagents that are mixed just prior to use.

For Research Use Only. Not for use in diagnostic procedures.
Especificaciones
Tipo de célulaMammalian Cells
Método de detecciónColorimétrico
Tipo de coloranteOtras etiquetas o colorantes
FormatoPlaca de 96 pocillos
Cantidad1000 assays
Condiciones de envíoTemperatura ambiente
Emission570
Para utilizar con (aplicación)Ensayo de viabilidad, Proliferation Assay, Cytotoxicity Assay
Para utilizar con (equipo)Lector de microplacas
Línea de productosCyQUANT™
Tipo de productoEnsayo de viabilidad celular
Unit Size1,000 assays
Contenido y almacenamiento
Almacenar a 2–8 °C y proteger de la luz.
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N.º de loteCertificate TypeDateCatalog Number(s)
3148415Certificate of Analysis20 jun 2025V13154
3148388Certificate of Analysis17 jun 2025X12223
3123587Certificate of Analysis04 may 2025X12223
3143140Certificate of Analysis22 abr 2025V13154
3143136Certificate of Analysis10 abr 2025V13154
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Preguntas frecuentes

This is true for MTT (as well as XTT, AlamarBlue Cell Viability Reagent, and PrestoBlue Cell Viability Reagent). CyQUANT Direct will also only count live cells, but for a different reason. The dye in it is a green-fluorescent nucleic acid stain, which will bind to DNA in all cells, live or dead, without the need for cellular metabolism. However, there is a non-cell-permeable quenching reagent in the kit which will both quench extracellular fluorescence (and thus this is a no-wash assay) AND will quench the fluorescence in dead cells (but not live cells).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

This is true for MTT (as well as XTT, AlamarBlue Cell Viability Reagent, and PrestoBlue Cell Viability Reagent). CyQUANT Direct will also only count live cells, but for a different reason. The dye is a green-fluorescent nucleic acid stain, which will bind to DNA in all cells, live or dead, without the need for cellular metabolism. However, there is a cell impermeant quenching reagent in the kit which will quench both extracellular fluorescence (and thus this is a no-wash assay) and the fluorescence in dead cells (but not live cells).

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

MTT is reduced only by live cells and the rate of dye reduction may vary from cell to cell based on their life cycle stage, age, health and other factors. This assay would not reveal the number of dead cells or a ratio of live to dead cells. A standard curve is recommended to determine an approximate number of live cells per sample.

To obtain a ratio of live to dead cells, use a two color fluorescent assay, such as LIVE/DEAD Viability/Cytotoxicity Kit (Cat. No. L3224) or differences in intensity of a single fluorescent color, using a flow cytometer and the LIVE/DEAD Fixable Kits.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Citations & References (286)

Citations & References
Abstract
Large-scale chemical dissection of mitochondrial function.
Authors:Wagner BK,Kitami T,Gilbert TJ,Peck D,Ramanathan A,Schreiber SL,Golub TR,Mootha VK
Journal:Nature biotechnology
PubMed ID:18297058
Mitochondrial oxidative phosphorylation (OXPHOS) is central to physiology and disease pathogenesis. To systematically investigate its activity and regulation, we performed a wide range of assays of OXPHOS physiology and nuclear and mitochondrial gene expression across 2490 chemical perturbations in muscle cells. Through mining of the resulting compendium, we discovered that: ... More
Authors:
Journal:
PubMed ID:10896672
Detection of low copy numbers of HPV DNA by fluorescent in situ hybridization combined with confocal microscopy as an alternative to in situ polymerase chain reaction.
Authors:Lizard G, Chignol MC, Souchier C, Roignot P, Chardonnet Y, Schmitt D
Journal:J Virol Methods
PubMed ID:9672129
'In genital lesions infected by human papillomavirus (HPV), histological criteria and HPV DNA typing are of prognostic value. Therefore, non-radioactive methods such as in situ hybridization are used extensively since they preserve the histological organization of the tissue, and allow the detection and characterization of HPV DNA. However, the sensitivity ... More
Identification of an amino acid residue in multidrug resistance protein 1 critical for conferring resistance to anthracyclines.
Authors:Zhang DW, Cole SP, Deeley RG
Journal:J Biol Chem
PubMed ID:11278596
'Murine multidrug resistance protein 1 (mrp1), unlike human MRP1, does not confer resistance to anthracyclines. Previously, we have shown that a human/murine hybrid protein containing amino acids 959-1187 of MRP1 can confer resistance to these drugs. We have now examined the functional characteristics of mutant proteins in which we have ... More
Oxidative stress induces intracellular accumulation of amyloid beta-protein (Abeta) in human neuroblastoma cells.
Authors:Misonou H, Morishima-Kawashima M, Ihara Y
Journal:Biochemistry
PubMed ID:10841777
'Several lines of evidence suggest that enhanced oxidative stress is involved in the pathogenesis and/or progression of Alzheimer''s disease (AD). Amyloid beta-protein (Abeta) that composes senile plaques, a major neuropathological hallmark of AD, is considered to have a causal role in AD. Thus, we have studied the effect of oxidative ... More
286 total citations

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