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Thiazolyl Blue  (Synonyms: MTT; Thiazolyl Blue Tetrazolium bromide; Methylthiazolyldiphenyl-tetrazolium bromide)

Cat. No.: HY-15924 Purity: 99.84%
COA Handling Instructions

Thiazolyl Blue (MTT) is a cell-permeable and positively charged tetrazolium dye that is used to detect reductive metabolism in cells. Thiazolyl Blue is taken up by cells through the plasma membrane and then reduced to formazan by intracellular NAD (P) H-oxidoreductases. Thiazolyl Blue is frequently used in colorimetric assays to measure cell proliferation, cytotoxicity, and apoptosis.

For research use only. We do not sell to patients.

Thiazolyl Blue Chemical Structure

Thiazolyl Blue Chemical Structure

CAS No. : 298-93-1

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Customer Review

Based on 98 publication(s) in Google Scholar

Top Publications Citing Use of Products

98 Publications Citing Use of MCE Thiazolyl Blue

  • Biological Activity

  • Purity & Documentation

  • References

  • Customer Review

Description

Thiazolyl Blue (MTT) is a cell-permeable and positively charged tetrazolium dye that is used to detect reductive metabolism in cells. Thiazolyl Blue is taken up by cells through the plasma membrane and then reduced to formazan by intracellular NAD (P) H-oxidoreductases. Thiazolyl Blue is frequently used in colorimetric assays to measure cell proliferation, cytotoxicity, and apoptosis[1].

In Vitro

1.Preparation of MTT working solution
Dissolve MTT by PBS to obtain 5 mg/mL of MTT.
2.Cell proliferation detection ( 96-well plates)
2.1Inoculated cells:Prepared single cell suspension to prepared with culture medium containing 10% FBS and inoculated into 96-well plate with 1000-10000 cells per well, volume of 100 uL per well.
2.2Incubated cells: 37℃. 5% CO2, Incubate for 24-72 h.
2.3Add 10 μL MTT in per well and incubate for 4 h, Discard the supernatant, For suspended cells, need centrifuge firstly.
2.4Add 100 μL DMSO, shock 10 min to dissolve the crystal completely
2.5Monitor the absorbance increase with an absorbance plate reader at OD = 562 nm.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

414.32

Formula

C18H16BrN5S

CAS No.
Appearance

Solid

Color

Light yellow to yellow

SMILES

CC1=C(C)N=C(N2N=C(C3=CC=CC=C3)N=[N+]2C4=CC=CC=C4)S1.[Br-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 1 years; -20°C, 6 months (sealed storage, away from moisture and light)

Solvent & Solubility
In Vitro: 

DMSO : 25 mg/mL (60.34 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

H2O : 0.88 mg/mL (2.12 mM; Need ultrasonic and warming)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.4136 mL 12.0680 mL 24.1359 mL
5 mM 0.4827 mL 2.4136 mL 4.8272 mL
View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 1 years; -20°C, 6 months (sealed storage, away from moisture and light). When stored at -80°C, please use it within 1 years. When stored at -20°C, please use it within 6 months.

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

  • Molarity Calculator

  • Dilution Calculator

Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

Mass
=
Concentration
×
Volume
×
Molecular Weight *

Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

Concentration (start)

C1

×
Volume (start)

V1

=
Concentration (final)

C2

×
Volume (final)

V2

In Vivo:

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

    Solubility: ≥ 2.08 mg/mL (5.02 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

    Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
  • Protocol 2

    Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

    Solubility: ≥ 2.08 mg/mL (5.02 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.8 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

    Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.

For the following dissolution methods, please prepare the working solution directly. It is recommended to prepare fresh solutions and use them promptly within a short period of time.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

  • Protocol 1

    Add each solvent one by one:  PBS

    Solubility: 6.67 mg/mL (16.10 mM); Clear solution; Need ultrasonic

In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

Dosage

mg/kg

Animal weight
(per animal)

g

Dosing volume
(per animal)

μL

Number of animals

Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO +
+
%
Tween-80 +
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration: mg/mL
Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

*In solvent : -80°C, 1 years; -20°C, 6 months (sealed storage, away from moisture and light)

The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
 If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Purity & Documentation

Purity: 99.84%

References

Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 1 years; -20°C, 6 months (sealed storage, away from moisture and light). When stored at -80°C, please use it within 1 years. When stored at -20°C, please use it within 6 months.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
H2O / DMSO 1 mM 2.4136 mL 12.0680 mL 24.1359 mL 60.3398 mL
DMSO 5 mM 0.4827 mL 2.4136 mL 4.8272 mL 12.0680 mL
10 mM 0.2414 mL 1.2068 mL 2.4136 mL 6.0340 mL
15 mM 0.1609 mL 0.8045 mL 1.6091 mL 4.0227 mL
20 mM 0.1207 mL 0.6034 mL 1.2068 mL 3.0170 mL
25 mM 0.0965 mL 0.4827 mL 0.9654 mL 2.4136 mL
30 mM 0.0805 mL 0.4023 mL 0.8045 mL 2.0113 mL
40 mM 0.0603 mL 0.3017 mL 0.6034 mL 1.5085 mL
50 mM 0.0483 mL 0.2414 mL 0.4827 mL 1.2068 mL
60 mM 0.0402 mL 0.2011 mL 0.4023 mL 1.0057 mL

* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.

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Thiazolyl Blue Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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