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  2. Fluorescent Dye Reactive Oxygen Species Mitochondrial Metabolism
  3. MitoSOX Red

MitoSOX Red is a live cell fluorescent probe that specifically targets mitochondria and is cell membrane permeable. MitoSOX Red enters mitochondria and is oxidized by superoxide but not by other ROS or RNS generating systems. The oxidized MitoSOX Red then binds to nucleic acids in mitochondria/nucleus, producing strong red fluorescence. MitoSOX Red can be used as a fluorescent indicator to specifically detect superoxide. In addition, superoxide dismutase (SOD) can prevent the oxidation of MitoSOX Red.
Excitation/emission wavelength: 510/580 nm.

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

MitoSOX Red Chemical Structure

MitoSOX Red Chemical Structure

CAS No. : 1003197-00-9

Size Price Stock Quantity
50 μg USD 62 In-stock
100 μg USD 100 In-stock
500 μg USD 340 In-stock

* Please select Quantity before adding items.

This product is a controlled substance and not for sale in your territory.

Customer Review

Based on 21 publication(s) in Google Scholar

Top Publications Citing Use of Products
  • Biological Activity

  • Purity & Documentation

  • References

  • Customer Review

Description

MitoSOX Red is a live cell fluorescent probe that specifically targets mitochondria and is cell membrane permeable. MitoSOX Red enters mitochondria and is oxidized by superoxide but not by other ROS or RNS generating systems. The oxidized MitoSOX Red then binds to nucleic acids in mitochondria/nucleus, producing strong red fluorescence. MitoSOX Red can be used as a fluorescent indicator to specifically detect superoxide. In addition, superoxide dismutase (SOD) can prevent the oxidation of MitoSOX Red.
Excitation/emission wavelength: 510/580 nm.

In Vitro

General Protocol

1. Preparation of MitoSOX Red working solution
1.1 Preparation of the stock solution
Dissolve 50 μg MitoSOX Red in 13 μL DMSO to obtain 5 mM of stock solution.
Note: It is recommended to store the stock solution at -20°C and -80°C away from light and avoid repetitive freeze-thaw cycles.
1.2 Preparation of MitoSOX Red working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of working solution.
Note: Please adjust the concentration of MitoSOX Red working solution according to the actual situation.
2. Cell staining
2.1 Suspension cells(6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL.
b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslip from the medium and aspirate excess medium.
c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 minutes.
d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

Storage
-20°C, 1 year. Protect from light

Precautions
1. Please adjust the concentration of MitoSOX Red working solution according to the actual situation.
2. This product is for R&D use only, not for drug, household, or other uses.
3. For your safety and health, please wear a lab coat and disposable gloves to operate.

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

Molecular Weight

759.70

Formula

C43H43IN3P

CAS No.
Appearance

Solid

Color

Yellow to brown

Emission (Em)

580

Excitation (Ex)

510

SMILES

NC1=CC(C(C2=CC=CC=C2)N(CCCCCC[P+](C3=CC=CC=C3)(C4=CC=CC=C4)C5=CC=CC=C5)C6=CC(N)=CC=C76)=C7C=C1.[I-]

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

-20°C, protect from light, stored under nitrogen

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

Solvent & Solubility
In Vitro: 

DMSO : 100 mg/mL (131.63 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 1.3163 mL 6.5815 mL 13.1631 mL
5 mM 0.2633 mL 1.3163 mL 2.6326 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 (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

  • 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: ≥ 2.5 mg/mL (3.29 mM); Clear solution

    This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).

    Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 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.
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, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)

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: 95.25%

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 (protect from light, stored under nitrogen). 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 1 mM 1.3163 mL 6.5815 mL 13.1631 mL 32.9077 mL
5 mM 0.2633 mL 1.3163 mL 2.6326 mL 6.5815 mL
10 mM 0.1316 mL 0.6582 mL 1.3163 mL 3.2908 mL
15 mM 0.0878 mL 0.4388 mL 0.8775 mL 2.1938 mL
20 mM 0.0658 mL 0.3291 mL 0.6582 mL 1.6454 mL
25 mM 0.0527 mL 0.2633 mL 0.5265 mL 1.3163 mL
30 mM 0.0439 mL 0.2194 mL 0.4388 mL 1.0969 mL
40 mM 0.0329 mL 0.1645 mL 0.3291 mL 0.8227 mL
50 mM 0.0263 mL 0.1316 mL 0.2633 mL 0.6582 mL
60 mM 0.0219 mL 0.1097 mL 0.2194 mL 0.5485 mL
80 mM 0.0165 mL 0.0823 mL 0.1645 mL 0.4113 mL
100 mM 0.0132 mL 0.0658 mL 0.1316 mL 0.3291 mL
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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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MitoSOX Red
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