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AAT Bioquest

OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor

Product key features

  • Readily used for live cell imaging 
  • Provide high selectivity for mitochondrial lipid peroxidation
  • The ratioable modality provides the highest accuracy
  • Suitable for complicated systems

Product description

A wide range of diseases is considered to result from mitochondrial oxidative damage that is associated with the lipid peroxidation of mitochondrial inner membranes. There are no specific methods to assess mitochondrial lipid peroxidation in live cells. To address this unmet need, AAT Bioquest has developed the OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor, a fluorescent mitochondria-targeted probe that detects lipid peroxidation in live cells. OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor enters cells rapidly, and selectively accumulates in mitochondria. It has a high specificity for the detection of mitochondrial lipid peroxidation. Mitochondrial lipid peroxidation results in a great change of fluorescence intensity ratio at 520 nm/590 nm, which can conveniently be monitored by fluorimetry, fluorescence microscopy or a fluorescence microplate reader. The fluorescence intensities are collected with FITC and Cy3/TRITC filter sets respectively at 520 nm/590 nm.

Example protocol

AT A GLANCE

Important Note

Before initial use, thaw OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor at room temperature and briefly centrifuge to collect the dried pellet.

Protocol Summary
  1. Prepare and treat cells as needed in growth medium

  2. Stain cells with OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor working solution

  3. Incubate samples at 37 °C in a 5% CO₂ incubator for 30–60 minutes

  4. Monitor fluorescence intensity with FITC and Cy3 filters

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Protect from light and avoid repeated freeze-thaw cycles

OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor Stock Solution
  1. Prepare 2 to 5 mM stock solution in DMSO. For example, add 25 μL of DMSO into one vial to create a 5 mM OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor stock solution.

    Note: Prepare single-use aliquots of the stock solution and store at ≤ -20°C. Protect from light and avoid repeated freeze-thaw cycles.

PREPARATION OF WORKING SOLUTION

OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor Working Solution
  1. Prepare a 500 to 1000 nM OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor working solution. For example, add 2 μL of 5 mM OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor stock solution to 10 mL of cell culture medium or HHBS buffer (AAT cat# 20011).

    Note: Protect the working solution from light by covering it with foil or placing it in the dark.

    Note: For best results, use the solution within 2 hours of its preparation.

    Note: 10 mL of working solution is enough for 100 tests.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Plate cells as needed in a 96-well black wall, clear bottom plate.

  2. Add treatment to induce lipid peroxidation.

  3. Add 100 µL of OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor working solution to cells.

  4. Incubate cells at 37 ºC in a 5% CO2 incubator for 30–60 minutes, protected from light.

    Note: The optimal concentration and incubation time may vary by cell line; we recommend testing with different concentrations.

  5. Optional: Remove the dye working solution and wash cells twice with HHBS buffer if background fluorescence is observed.

  6. Add HHBS buffer and analyze the cells using a fluorescence microscope with a FITC and Cy3 filter set.

References

View all 50 references: Citation Explorer
A Metastable Semiquinone Molecular Switch Modulated by Ascorbate/O2: A Study from a System Far-From-Equilibrium to Biological Assays in Mitochondria.
Authors: Rodríguez-Valdez, Gabriela and Martínez-Cerda, Marlen E and Mejía-Reyes, Jisell G and Tapia-Juárez, Melissa and Olmos-Orizaba, Eridani and Cortés-Rojo, Christian and Cortés-García, Carlos J and Contreras-Celedón, Claudia A and Solorio-Alvarado, Cesar R and Chacón-García, Luis
Journal: Chembiochem : a European journal of chemical biology (2024): e202400401
Synergy between nanoplastics and benzo[a]pyrene promotes senescence by aggravating ferroptosis and impairing mitochondria integrity in Caenorhabditis elegans.
Authors: Ren, Huasheng and Yin, Kai and Lu, Xinhe and Liu, Jiaojiao and Li, Dandan and Liu, Zuojun and Zhou, Hailong and Xu, Shunqing and Li, Hanzeng
Journal: The Science of the total environment (2024): 174418
Excitotoxicity, Oxytosis/Ferroptosis, and Neurodegeneration: Emerging Insights into Mitochondrial Mechanisms.
Authors: Khan, Sameera and Bano, Nargis and Ahamad, Shakir and John, Urmilla and Dar, Nawab John and Bhat, Shahnawaz Ali
Journal: Aging and disease (2024)
Ferroptosis Integrates Mitochondrial Derangements and Pathological Inflammation to Promote Pulmonary Hypertension.
Authors: Kazmirczak, Felipe and Vogel, Neal T and Prisco, Sasha Z and Patterson, Michael T and Annis, Jeffrey and Moon, Ryan T and Hartweck, Lynn M and Mendelson, Jenna B and Kim, Minwoo and Mancipe, Natalia Calixto and Markowski, Todd and Higgins, LeAnn and Guerrero, Candace and Kremer, Ben and Blake, Madelyn L and Rhodes, Christopher J and Williams, Jesse W and Brittain, Evan L and Prins, Kurt W
Journal: bioRxiv : the preprint server for biology (2024)
Cardiotoxicity of Iron and Zinc and Their Association with the Mitochondrial Unfolded Protein Response in Humans.
Authors: Mirosevic, Vid and Svagusa, Tomo and Matic, Natalija and Maldini, Kresimir and Siljeg, Mario and Milicic, Davor and Gasparovic, Hrvoje and Rudez, Igor and Sepac, Ana and Gojmerac, Lucija and Kulic, Ana and Bakovic, Petra and Sedlic, Filip
Journal: International journal of molecular sciences (2024)
Page updated on December 13, 2024

Ordering information

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Catalog Number21511
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Additional ordering information

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Physical properties

Molecular weight

716.02

Solvent

DMSO

Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Freeze (< -15 °C); Minimize light exposure

Platform

Fluorescence microscope

ExcitationFITC and Cy3, TRITC Filter
EmissionFITC and Cy3, TRITC Filter
Recommended plateBlack wall, clear bottom
Fluorescence response of OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor (1000 nM) in HeLa cells with or without Erastin (10 µM) treatment at 37 ºC in a 5% CO2 incubator for 30 minutes. The fluorescence intensities were monitored with fluorescence microscopy using FITC and Cy3 filters.
Fluorescence response of OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor (1000 nM) in HeLa cells with or without Erastin (10 µM) treatment at 37 ºC in a 5% CO2 incubator for 30 minutes. The fluorescence intensities were monitored with fluorescence microscopy using FITC and Cy3 filters.
Fluorescence response of OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor (1000 nM) in HeLa cells with or without Erastin (10 µM) treatment at 37 ºC in a 5% CO2 incubator for 30 minutes. The fluorescence intensities were monitored with fluorescence microscopy using FITC and Cy3 filters.