298-93-1

Basic information

• Product Name: Thiazolyl Blue
• Synonyms: METHYLTHIAZOLYLDIPHENYL-TETRAZOLIUM BROMIDE;METHYLTHIAZOLETETRAZOLIUM;3-(4,5-DIMETHYL-2-THIAZOL-2-YL)-2,5-DIPHENYL-2H-TETRAZOLIUM BROMIDE;3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHYL-2 H-TETRAZOLIUM BROMIDE;3,[4,5-DIMETHYLTHIAZOL-2-YL]-2,5-DIPHENYLTETRAZOLIUM BROMIDE;3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHENYL-2H-TETRAZOLIUM BROMIDE;3-(4,5-DIMETHYLITHIAZOLYL-2)-2,5-DIPHENYL MONOTETRAZOLIUM BROMIDE;3-(4,5-DIMETHYL-2-THIAZOLYL)-2,5-DIPHENYLTETRAZOLIUM BROMIDE
• CAS NO: 298-93-1
• Molecular Formula: Br*C₁₈H₁₆N₅S
• Molecular Weight: 414.32
• EINECS: 206-069-5
• Product Categories: Tetrazolium Salts;Tetrazolium Salts & Formazans;Chemistry;marker;Tetrazolium salt
• Mol File: 298-93-1.mol
• Article Data: 0

Chemical Properties

• Melting Point: 195 °C (dec.)(lit.)
• Appearance: Yellow to orange/powder
• Density: 1.4886 (estimate)
• Storage Temp.: 2-8°C
• Solubility: methanol: 5 mg/mL, clear
• Water Solubility: Slightly soluble
• Sensitive: Light Sensitive
• Stability: Hygroscopic
• BRN: 3825277
• CAS DataBase Reference: Thiazolyl Blue(CAS DataBase Reference)
• NIST Chemistry Reference: Thiazolyl Blue(298-93-1)
• EPA Substance Registry System: Thiazolyl Blue(298-93-1)

Safety Data

• Hazard Codes: Xi,T,Xn
• Statements: 46-68-36/37/38
• Safety Statements: 22-24/25-53-45-36/37-26
• WGK Germany: 3
• RTECS: XF8060000
• F: 3-8-10
• TSCA: Yes
• Hazardous Substances Data: 298-93-1(Hazardous Substances Data)

Usage

Description

Thiazolyl Blue, also known as MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), is a cell-permeable and positively charged colorimetric agent. It is widely used to detect reductive metabolism in cells and to measure cell proliferation, cytotoxicity, and apoptosis. Thiazolyl Blue is a direct indicator of cytotoxicity and is utilized for studying nicotinamide adenine dinucleotide phosphate (NADP)-dependent dehydrogenases. It is also known as Tetrazolium bromide, Methylthiazolyldiphenyl-tetrazolium bromide, and Methylthialazole Tetrazolium. As a dye and colorimetric reagent, Thiazolyl Blue plays a significant role in various biological and medical applications.

Uses

Used in Cell Proliferation Measurement:
Thiazolyl Blue is used as a colorimetric reagent for measuring cell proliferation. It 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.
Used in Cytotoxicity and Apoptosis Detection:
Thiazolyl Blue is used as a direct indicator of cytotoxicity, which is essential for screening cancer drugs. It is also utilized in detecting apoptosis, a process of programmed cell death, providing valuable insights into cellular responses to various treatments.
Used in Nicotinamide Adenine Dinucleotide Phosphate (NADP)-Dependent Dehydrogenase Studies:
As an electron acceptor, Thiazolyl Blue is employed for studying NADP-dependent dehydrogenases, which are crucial enzymes involved in cellular metabolism and energy production.
Used in Histochemical/Cytochemical Applications:
Thiazolyl Blue has been used as a histochemical and cytochemical reagent, allowing for the visualization and analysis of cellular structures and functions.
Used in Detection of NAD and ADP-Linked Enzyme Systems:
Thiazolyl Blue is used for the detection of NAD (Nicotinamide adenine dinucleotide) and ADP-linked enzyme systems, which are essential for cellular energy metabolism and various biological processes. However, it cannot be used to detect ADP-linked enzyme systems in tissue due to the binding of the cation by the cyanide trap used.
Used in Oxidative Systems:
The formazan produced from the reduction of Thiazolyl Blue can chelate with metals such as nickel, copper, and cobalt. The cobalt chelate has been used in oxidative systems, providing further applications in various research and industrial settings.

Biochem/physiol Actions

Cell permeable: yes

Purification Methods

It is recrystallised by dissolving in MeOH containing a few drops of HBr and then adding dry Et2O to complete the crystallisation, wash the needles with Et2O and dry them in a vacuum desiccator over KOH. [Beyer & Pyl Chem Ber 87 1505 1954, Beilstein 27 III/IV 6045.]

InChI:InChI=1/C18H17N5S.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,(H,20,21);1H

Synthetic route

N-<4,5-dimethyl-thiazol-2-yl>-3,N'''-diphenyl-formazan → thiazolyl blue (298-93-1)

Conditions	Yield
With N-Bromosuccinimide; ethyl acetate

sodium molybdate dihydrate (7631-95-0) + 1,2-dihydroxy-4-nitrobenzene (3316-09-4) + thiazolyl blue (298-93-1) → [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium][MoO2(4-nitrocatechol(-2H))2]

Conditions	Yield
In water mixing of aq. solns. of Na2MoO4*2H2O, 4-nitrocatechol and 3-(4,5-dimethylthiaozol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide in acidic medium (pH2.8 - 4.0); pptn., filtration, washing with cold water, drying at 75-80°C;

thiazolyl blue (298-93-1) → 1-(4,5-dimethyl-2-thiazoyl)-3,5-diphenylformazan (23305-68-2)

Conditions	Yield
With dehydrogenase
With succinate dehydrogenase at 37℃; for 4h;

iron(III) ammonium sulfate dodecahydrate + 1,2-dihydroxy-4-nitrobenzene (3316-09-4) + thiazolyl blue (298-93-1) → Fe(3+)*3C6H3NO4(2-)*3C18H16N5S(1+)

Conditions	Yield
With sulfuric acid In water

ammonium metavanadate + thiazolyl blue (298-93-1) + 5-methyl-4-(2-thiazolylazo)resorcinol (37422-56-3) → 2O2V(1+)*2C10H7N3O2S(2-)*2C18H16N5S(1+)

Conditions	Yield
In chloroform; water

thiazolyl blue (298-93-1) → (E,Z)-5-(4,5-dimethylthiazol-2-yl)-1,3-diphenylformazan

Conditions	Yield
With mitochondrial reductase Enzymatic reaction;

thiazolyl blue (298-93-1) → MTT formazan metabolite

Conditions	Yield
With A549 cells for 1h; Enzymatic reaction;
With mitochondrial dehydrogenase Enzymatic reaction;

Downstream Products

• 23305-68-2 1-(4,5-dimethyl-2-thiazoyl)-3,5-diphenylformazan