At physiological pH, phenol red strongly absorbs in the same wavelength range as the MTT assay’s readout product. Washing cells thoroughly to remove the media and phenol red minimizes this interference.
M5655
Azul de tiazolil Bromuro de tetrazolio
powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC)
Sinónimos:
Bromuro de 3-(4,5-dimetil-2-tiazolil)-2,5-difenil-2H-tetrazolio, Bromuro de metiltiazolildifenil-tetrazolio, MTT
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Fórmula empírica (notación de Hill):
C18H16BrN5S
Número CAS:
Peso molecular:
414.32
UNSPSC Code:
12352207
NACRES:
NA.75
PubChem Substance ID:
EC Number:
206-069-5
Beilstein/REAXYS Number:
4081397
MDL number:
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Servicio técnico
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Permítanos ayudarleproduct line
BioReagent
Quality Segment
assay
≥97.5% (HPLC)
form
powder
technique(s)
cell culture | insect: suitable, cell culture | mammalian: suitable
mp
195 °C (dec.) (lit.)
solubility
H2O: 5 mg/mL
storage temp.
2-8°C
SMILES string
[Br-].Cc1nc(sc1C)-[n+]2nc(nn2-c3ccccc3)-c4ccccc4
InChI
1S/C18H16N5S.BrH/c1-13-14(2)24-18(19-13)23-21-17(15-9-5-3-6-10-15)20-22(23)16-11-7-4-8-12-16;/h3-12H,1-2H3;1H/q+1;/p-1
InChI key
AZKSAVLVSZKNRD-UHFFFAOYSA-M
Application
El bromuro de tetrazolio de azul de tiazolilo se ha utilizado como indicador colorimétrico de la actividad metabólica en análisis de viabilidad celular (análisis MTT). Se ha utilizado también para determinar la proliferación celular.
El azul de tiazolil Azul tetrazolio (MTT) puede utilizarse para medir la proliferación celular. El MTT produce una disolución amarillenta que vira a formazano de MTT azul violeta insoluble en agua por acción de las deshidrogenasas mitocondriales de las células vivas. Los cristales azules se solubilizan con isopropanol acidificado y la intensidad se mide colorimétricamente a 570 nm.
El azul de tiazolil Azul tetrazolio (MTT) puede utilizarse para medir la proliferación celular. El MTT produce una disolución amarillenta que vira a formazano de MTT azul violeta insoluble en agua por acción de las deshidrogenasas mitocondriales de las células vivas. Los cristales azules se solubilizan con isopropanol acidificado y la intensidad se mide colorimétricamente a 570 nm. El MTT se ha utilizado como un reactivo histoquímico o citoquímico y para la detección del NAD. Los sistemas enzimáticos ligados a ADP de los tejidos no pueden detectarse con MTT, debido a la unión del catión por la trampa de cianuro utilizada. El MTT se reduce rápidamente al formazano, que forma complejos con el níquel, el cobre y el cobalto; los quelatos de cobalto se han utilizado en los sistemas oxidativos.
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Este artículo | |||
|---|---|---|---|
| description powder, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥97.5% (HPLC) | description 98% | description suitable for microbiology, ≥90% (T) | description used in colorimetric determination of reducing compounds |
| technique(s) cell culture | insect: suitable, cell culture | mammalian: suitable | technique(s) titration: suitable | technique(s) - | technique(s) - |
| solubility H2O: 5 mg/mL | solubility ethanol: soluble 20 mg/mL, H2O: 5 mg/mL, 2-methoxyethanol: soluble 20 mg/mL | solubility methanol: 0.5 g/10 mL ( hot) | solubility methanol: 50 mg/mL, clear |
| form powder | form powder | form powder | form powder |
| assay ≥97.5% (HPLC) | assay 98% | assay ≥90% (T) | assay - |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. - | storage temp. room temp |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
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Palabra de advertencia
Warning
Códigos de peligro
Indicaciones de precaución
Hazard Classifications
Eye Irrit. 2 - Muta. 2 - Skin Irrit. 2 - STOT SE 3
Órganos diana
Respiratory system
Clase de almacenamiento
11 - Combustible Solids
¿Qué está pasando?
WGK 3
Punto de inflamación (°F)
Not applicable
Punto de inflamación (°C)
Not applicable
EPI (equipos de protección individual)
Eyeshields, Gloves, type N95 (US)
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Why can't a medium with phenol red be used? Is it because it interferes with MTT assay results? If after incubation with MTT, cells are washed with PBS and DMSO is used to dissolve the crystals, does this still apply?
1 answer-
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Should I prepare fresh for each use or I directly resuspend in 20 ml of water (for the 100 mg bottle) and store? In the latter case, how can I store the solution?
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The solution may be filtered through a 0.2 μm filter and stored at 2-8°C for frequent use or stored in aliquots and frozen at -20°C for extended periods.
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What's the difference between M5655 and M2003?
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Product M5655 is Thiazolyl Blue Tetrazolium Bromide and Product M2003 is Thiazolyl blue formazan.
MTT (Cat. no. M5655) is used in the measurement of cell proliferation. MTT produces a yellowish solution that is converted to dark blue, water-insoluble MTT formazan (Product M2003) by mitochondrial dehydrogenases of living cells. The blue MTT formazan crystals are solubilized with acidified isopropanol and the intensity is measured colorimetrically at 570 nm. Thus, the amount of blue formazan is proportional to the number of viable cells. For more information regarding this particular assay, we do offer an assay kit, TOX1, which is based on this principle: https://www.sigmaaldrich.com/product/sigma/tox1.
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What is the Department of Transportation shipping information for this product?
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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What type of multi-well plate should be used for the Thiazolyl Blue Tetrazolium Bromide (MTT) assay?
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It is recommended that flat bottom multi-well plates be used for the MTT assay. The use of tissue culture treated plates is only important if necessary for good cell growth. If cells are grown in larger plates, the supernatant after cell lysis can be transferred to a 96 well plate for reading.
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What can be used to solubilize MTT formazan (the product of the Thiazolyl Blue Tetrazolium Bromide - MTT reaction)?
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Mitochondrial dehydrogenases of viable cells cleave the tetrazolium ring of the MTT, yielding purple MTT formazan crystals. These crystals are not soluble in aqueous solutions; they may be dissolved in acidified isopropanol (0.1 N HCl in anhydrous isopropanol). If this does not lyse the cells, Triton X-100 can be added to the acidified isopropanol. A typical working concentration is 10% Triton X-100 (i.e., 1 volume of 100% Triton X-100 + 9 volumes of acidified isopropanol).
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Why is a reference wavelength used for data analysis of Thiazolyl Blue Tetrazolium Bromide (MTT)?
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The reference wavelength (630-690 nm) is used to adjust for the optical variation in the wells of the plate. It also corrects for dust and fingerprints that can be present. The assay is read at 570 nm. The reference wavelength needs to be far enough away from the assay wavelength so as not to affect those values.
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How can I store solutions of Thiazolyl Blue Tetrazolium Bromide (MTT)?
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Reconstituted MTT solution is stable for at least 6 months when stored frozen (-20 °C). Storage of reconstituted MTT solution at 2-8°C for more than 2 weeks may cause decomposition and yield erroneous results.
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What can be used to make a solution of Thiazolyl Blue Tetrazolium Bromide (MTT)?
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We test the solubility of MTT in water at a concentration of 5 mg/mL. For the MTT viability assay, we dissolve Product No. M5655 at a concentration of 5 mg/ml in RPMI-1640 without phenol red. This medium is available as a powder (Product No.R8755) or liquid (Product No. R7509). MTT can be solubilized in any culture media or balanced salt solution that does not contain phenol red as a pH indicator.
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How can Thiazolyl Blue Tetrazolium Bromide be used for staining dehydrogenase in tissue sections?
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Products M2128 or M5655, Thiazolyl Blue Tetrazolium Bromide can be used for staining dehydrogenase in tissue sections. See the method below. Reagent PreparationMTT (2 mg/ml distilled water) 2.5 ml0.2 M Tris buffer, pH 7.4 2.5 ml0.5 M cobalt chloride 0.5 ml0.05 M magnesium chloride 1.0 mlDistilled water 2.5 mlThe pH of this solution is adjusted to 7.0 if necessary. Aliquot and store at -20°C until required. Sample PreparationCut cytostat sections at 5 to 7 microns. Do not fix. Method:1. Incubate sections in appropriate incubation solution at 37°C for 30-60 minutes.2. Transfer sections to 15% formol saline, 15 minutes.3. Wash in distilled water4. Counterstain in 2% methyl green if required.5. Wash in distilled water.6. Mount in glycerin Jelly. ResultsEnzyme: Black formazan deposits with MTT Source: Bancroft and Gamble, Theory and Practice of Histological Techniques, Fifth Edition. Churchill Livingstone, London, 2002, page 610.
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