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  4. Tetrazolium Red

Tetrazolium Red  (Synonyms: 2,3,5-Triphenyltetrazolium chloride; TPTZ; TTC)

Cat. No.: HY-D0714 Purity: 99.85%
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Tetrazolium Red (2,3,5-Triphenyltetrazolium chloride; TTC) is a not brain-penetrant, colorless, water-soluble dye that is reduced by mitochondrial enzymes to a deep red, water-insoluble compound (formazan) mainly in the mitochondria of living cells. Tetrazolium Red is used to observe the activity of dehydrogenase, and it turns colorless to red when exposed to hydrogen. Tetrazolium Red distinguishes between surviving and infarcted brain tissue after stroke. Tetrazolium Red has been used to stain heart tissue to measure the extent of acute lesions and also used to stain brain tissue to detect the size of the infarcted area. The absorption wavelength of Tetrazolium Red is 570 nm.

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

Tetrazolium Red Chemical Structure

Tetrazolium Red Chemical Structure

CAS No. : 298-96-4

Size Price Stock Quantity
Solid + Solvent (Highly Recommended)
10 mM * 1 mL in DMSO
ready for reconstitution
 USD 28 In-stock
Solution
10 mM * 1 mL in DMSO USD 28 In-stock
Solid
500 mg USD 25 In-stock
5 g USD 39 In-stock
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Based on 3 publication(s) in Google Scholar

Tetrazolium Red Related Antibodies

Top Publications Citing Use of Products

  • Biological Activity

  • Purity & Documentation

  • References

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Description

Tetrazolium Red (2,3,5-Triphenyltetrazolium chloride; TTC) is a not brain-penetrant, colorless, water-soluble dye that is reduced by mitochondrial enzymes to a deep red, water-insoluble compound (formazan) mainly in the mitochondria of living cells. Tetrazolium Red is used to observe the activity of dehydrogenase, and it turns colorless to red when exposed to hydrogen. Tetrazolium Red distinguishes between surviving and infarcted brain tissue after stroke. Tetrazolium Red has been used to stain heart tissue to measure the extent of acute lesions and also used to stain brain tissue to detect the size of the infarcted area. The absorption wavelength of Tetrazolium Red is 570 nm[1][2][3][4].

In Vitro

Tetrazolium Red staining of the size of the ischemic area[3]:
 1) Preparation of human umbilical mesenchymal stem cells (HUMSCs)
HUMSCs are dissociated from the infants and cultured by 10% fetal bovine serum in DMEM at 37℃.
2) Animal Surgery
The rats are are anesthetized with chloral hydrate (400 mg/kg, i.p.) and are performed MCAO surgery and reperfusion.
NOTE: MCAO surgery, a hole was drilled by the right orbit to expose the right middle cerebral artery. The artery was ligated at the same time as both common carotid arteries were clamped simultaneously for 90 minutes, after which the ligature and clamping clips were removed to restore blood flow.
3) Transplantation of HUMSCs
A total of 5x105 HUMSCs are grafted into the infarct cortex of each rat 24 hours after MCAO by 2 injections.
4) Infarct Cortex Identification
Rats are deeply anesthetized and decapitated. Coronal sections of the brains were sliced at 2 mm, immersed in 2% Tetrazolium Red, and then fixed with 10% formalin.
5) Data analysis (digital images)
The size of the infarct area, which is devoid of red staining is determined on the digital images using ImagePro software.
Tetrazolium Red staining of cell proliferation and viability measurement[4]:
 1) Preparation: 2-5 mg/mL Tetrazolium Red in PBS.
2) Isolated cells are counted manually with a hemocytometer and then various concentrations (5x104 to 1x106 are plated into 24-well culture plates, fed 1 mL of control culture media and returned to the incubator for a 24 h period to allow for cell to reattach to the plastic culture plates.
3) The next day, media is removed and cells are fed 1 mL fresh media, and then 200 mL of 5 mg/mL Tetrazolium Red is added to each well so that final concentration is 0.83 mg/mL Tetrazolium Red. Plates are returned to the incubator (37℃) for a 4 h incubation period.
4) Media is removed and 1 mL of isopropanol is added to each well to dissolve the formazan crystals. Optical density is measured at 570 nm.

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

Tetrazolium Red Related Antibodies
In Vivo

Tetrazolium Red staining of brain damage and the size of the infarct area[1]：
 1)For lipopolysaccharide (LPS)-sensititized neonatal cerebral hypoxia-ischemia (HI), LPS (0.3 mg/kg) is i.p.-injected to 5-day-old CD1 mice at 72 h before HI. At postnatal day 8, mice are anesthetized by 2% isoflurane and subjected to permanent ligation of the right common carotid artery. Mice recover for 1.5 h and are then exposed to hypoxia in glass chambers containing 10% oxygen and 90% N2 in a waterbacth kept at 37℃. After hypoxic exposure, mice are returned to dams in the animal care facility. The in-vivo Tetrazolium Red-labeling procedure is performed at 48 h recovery.
2)Mannitol (0.5 M-1.4M) prepared in PBS at a temperature of 37℃ is IP-injected to animals (~0.1 mL/g body weight) for 5 to 180 min to disrupt blood-brain-barrier (BBB). Mice are anesthetized with avertin and transcardially perfused of PBS followed by 10 mL of 2% Tetrazolium Red. At 10 min after transcardial Tetrazolium Red perfusion, the brains of animals are removed and placed into 4% paraformaldehyde. Alternatively, the extracted brains can be incubated in warm phosphate-saline buffer (PBS) for 30 min to enhance Tetrazolium Red-staining. Brains are removed and divided into the contralateral (the left cortex) and lesion sides (the right cortex) for protein extraction.
3)Data analysis (digital images): Brain damage is expressed as the ratio of the infarcted area (white area) to the area of the undamged, contralateral hemisphere.

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

334.80

Formula

C19H15ClN4
CAS No.
Appearance

Solid

Color

White to light yellow

SMILES

C1(C2=CC=CC=C2)=NN(C3=CC=CC=C3)[N+](C4=CC=CC=C4)=N1.[Cl-]
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

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

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

Solvent & Solubility
In Vitro: 

H2O : 50 mg/mL (149.34 mM; Need ultrasonic)

DMSO : 16.67 mg/mL (49.79 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.9869 mL 14.9343 mL 29.8686 mL
5 mM 0.5974 mL 2.9869 mL 5.9737 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, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

* 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: ≥ 1.67 mg/mL (4.99 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.67 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (16.7 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: ≥ 1.67 mg/mL (4.99 mM); Clear solution

    This protocol yields a clear solution of ≥ 1.67 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (16.7 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: 25 mg/mL (74.67 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.
Calculation results:
Working solution concentration: mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
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).
Purity & Documentation

Purity: 99.85%

SDS (544 KB)

COA (258 KB) LCMS (264 KB)

Handling Instructions (2659 KB)
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, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
DMSO / H2O 1 mM 2.9869 mL 14.9343 mL 29.8686 mL 74.6714 mL
5 mM 0.5974 mL 2.9869 mL 5.9737 mL 14.9343 mL
10 mM 0.2987 mL 1.4934 mL 2.9869 mL 7.4671 mL
15 mM 0.1991 mL 0.9956 mL 1.9912 mL 4.9781 mL
20 mM 0.1493 mL 0.7467 mL 1.4934 mL 3.7336 mL
25 mM 0.1195 mL 0.5974 mL 1.1947 mL 2.9869 mL
30 mM 0.0996 mL 0.4978 mL 0.9956 mL 2.4890 mL
40 mM 0.0747 mL 0.3734 mL 0.7467 mL 1.8668 mL
H2O 50 mM 0.0597 mL 0.2987 mL 0.5974 mL 1.4934 mL
60 mM 0.0498 mL 0.2489 mL 0.4978 mL 1.2445 mL
80 mM 0.0373 mL 0.1867 mL 0.3734 mL 0.9334 mL
100 mM 0.0299 mL 0.1493 mL 0.2987 mL 0.7467 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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Tetrazolium Red 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.

Keywords:

Tetrazolium Red298-96-42,3,5-Triphenyltetrazolium TPTZ TTCBiochemical Assay Reagentsdehydrogenasewater-solubleinfarctedstrokebrain tissueInhibitorinhibitorinhibit

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