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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
Before initial use, thaw OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor at room temperature and briefly centrifuge to collect the dried pellet.
Prepare and treat cells as needed in growth medium
Stain cells with OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor working solution
Incubate samples at 37 °C in a 5% CO₂ incubator for 30–60 minutes
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
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
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
Plate cells as needed in a 96-well black wall, clear bottom plate.
Add treatment to induce lipid peroxidation.
Add 100 µL of OxiVision™ Ratiometric Mitochondrial Lipid Peroxidation Sensor working solution to cells.
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.
Optional: Remove the dye working solution and wash cells twice with HHBS buffer if background fluorescence is observed.
Add HHBS buffer and analyze the cells using a fluorescence microscope with a FITC and Cy3 filter set.
References
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
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
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)
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)
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)
Safety data sheet