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M2128

Sigma-Aldrich

Thiazolyl Blue Tetrazolium Bromide

98%, powder

Synonym(s):

3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide, MTT, Methylthiazolyldiphenyl-tetrazolium bromide

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100 MG
$77.30
250 MG
$126.00
500 MG
$231.00
1 G
$384.00
5 G
$1,450.00
10 G
$2,610.00

$77.30

Availability

Estimated to ship on06 January 2025Details

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Select a Size

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100 MG
$77.30
250 MG
$126.00
500 MG
$231.00
1 G
$384.00
5 G
$1,450.00
10 G
$2,610.00

About This Item

Empirical Formula (Hill Notation):
C18H16BrN5S
CAS Number:
298-93-1
Molecular Weight:
414.32
Beilstein:
4081397
EC Number:
206-069-5
MDL number:
MFCD00011964
UNSPSC Code:
12352111
PubChem Substance ID:
24896603
NACRES:
NA.47

$77.30

Availability

Estimated to ship on06 January 2025Details

Request a Bulk Order

product name

Thiazolyl Blue Tetrazolium Bromide, 98%

Quality Level

200

Assay

98%

form

powder

technique(s)

titration: suitable

color

dark yellow

mp

195 °C (dec.) (lit.)

solubility

2-methoxyethanol: soluble 20 mg/mL
ethanol: soluble 20 mg/mL
H2O: 5 mg/mL

application(s)

diagnostic assay manufacturing
hematology
histology

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

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Application

Thiazolyl Blue Tetrazolium Blue (MTT) may be used in measurement of cell proliferation. MTT produces a yellowish solution that is converted to dark blue, water-insoluble MTT formazan by mitochondrial dehydrogenases of living cells. The blue crystals are solubilized with acidified isopropanol and the intensity is measured colorimetrically at 570 nm. MTT has been used as a histochemical/cytochemical reagent and for the detection of NAD. ADP-linked enzyme systems in tissue cannot be detected with MTT, due to binding of the cation by the cyanide trap used. MTT is rapidly reduced to the formazan, which chelates with nickel, copper, and cobalt; the cobalt chelate has been used in oxidative systems.
Thiazolyl Blue Tetrazolium Bromide has been used for the following assays:
  • Proliferation assay.[1][2]
  • TNF (tumor necrosis factor) bioassay.[3]

Pictograms

Health hazardExclamation mark
GHS08,GHS07

Signal Word

Warning

Hazard Statements

H315,H319,H335,H341

Hazard Classifications

Eye Irrit. 2 - Muta. 2 - Skin Irrit. 2 - STOT SE 3

Target Organs

Respiratory system

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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Protocols

Recycling Micro-Assay of β-NAD and β-NADH

Objective: To standardize a procedure for the recycling micro-assay of β-NAD and β-NADH

Questions

1–10 of 10 Questions  

  1. I used 0.05g in 10ml then syringe it and add 40ml. My question is how to calculate the percentage of viable cells? I used DMSO after MTT solution

    1 answer

    1. MTT is typically used for the semi-quantitative colorimetric measurement of cell viability. This analysis is performed in a multi-well format which screens for the viability of any given compound at any given concentration. The percentage of viable cells cannot be determined using this method. After adding a stop reagent, in this case DMSO, the formazan crystals will be dissolved, making manual counting a challenge, at best. When used in a multi-well plate format, cells are incubated, a stop solution is added, and the plate is read spectrophotometrically at 570 nM. Please see the link below to review the product datasheet:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/305/017/m2128pis.pdf

      For additional information, see the link below to the MTT kit protocol for product TOX1, a cytotoxicity kit that utilized MTT:
      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/333/177/tox1bul.pdf

      To measure cell viability as a percentage of an entire population, microscopic methods may be of interest. Reagents such as Trypan Blue allow for the clear identification of viable and non-viable cells. To review more information regarding this method, please see the link below:
      https://www.sigmaaldrich.com/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/mammalian-cell-culture/cell-quantification

      Helpful?

  2. What is the recommendation volume of 5 mg/ml of thiazolyl blue tetrazolium blue (in uL) per well to use for MTT assay? 

    1 answer

    1. Instructions for use are not provided for this product. However, there is an in vitro toxicology assay kit MTT based available. The catalog number is TOX1-1KT. In the kit, 15 mg of solid MTT is supplied. This is resuspended with 3 ml of media or a balanced salt solution without phenol red or serum. The link shared below will list the complete procedure

      https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/333/177/tox1bul.pdf

      Helpful?

  3. Is the MTT assay quantitative?

    1 answer

    1. The MTT asay is a method to assess cell viability.  This assay is a semiquantitative assay. The assay is used to compare the viability changes in treated cells to untreated cells. The absorbance is indicative of the cell number. The higher the absorbance, the greater the number of viable cells present. Most researchers compare the absorbance of the two samples as a ratio (ABS treated cells/ABS untreated cells) to get a fold increase/decrease in cell number.

      Helpful?

  4. What type of multi-well plate should be used for the Thiazolyl Blue Tetrazolium Bromide (MTT) assay?

    1 answer

    1. 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.

      Helpful?

  5. What can be used to solubilize MTT formazan (the product of the Thiazolyl Blue Tetrazolium Bromide - MTT reaction)?

    1 answer

    1. 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).

      Helpful?

  6. What can be used to make a solution of Thiazolyl Blue Tetrazolium Bromide (MTT)?

    1 answer

    1. 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.

      Helpful?

  7. What is the Department of Transportation shipping information for this product?

    1 answer

    1. 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.

      Helpful?

  8. How can I store solutions of Thiazolyl Blue Tetrazolium Bromide (MTT)?

    1 answer

    1. 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.

      Helpful?

  9. Why is a reference wavelength used for data analysis of Thiazolyl Blue Tetrazolium Bromide (MTT)?

    1 answer

    1. 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.

      Helpful?

  10. How can Thiazolyl Blue Tetrazolium Bromide be used for staining dehydrogenase in tissue sections?

    1 answer

    1. 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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