MitoSpy™ Red CMXRos

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Other Names
Mitochondrial labeling
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MitoSpyRedCMXRos_1_IF_121616
NIH3T3 cells were stained with 100 nM of MitoSpy™ Red CMXRos (red) for 20 minutes at 37°C, fixed with 1% paraformaldehyde (PFA) for ten minutes at room temperature, and permeabilized with 1X True Nuclear™ Perm Buffer for ten minutes at room temperature. Then the cells were stained with Flash Phalloidin™ NIR 647 (green) for 20 minutes at room temperature and counterstained with DAPI (blue). The image was captured with a 60x objective.
  • MitoSpyRedCMXRos_1_IF_121616
    NIH3T3 cells were stained with 100 nM of MitoSpy™ Red CMXRos (red) for 20 minutes at 37°C, fixed with 1% paraformaldehyde (PFA) for ten minutes at room temperature, and permeabilized with 1X True Nuclear™ Perm Buffer for ten minutes at room temperature. Then the cells were stained with Flash Phalloidin™ NIR 647 (green) for 20 minutes at room temperature and counterstained with DAPI (blue). The image was captured with a 60x objective.
  • MitoSpyRedCMXRos_2_FC_121616
    Human T-cell leukemia cell line, Jurkat, was treated with 1.0 µg/mL of LEAF™ purified anti-human CD95. At the end of incubation, cells were washed with PBS twice and stained with 25 nM of MitoSpy™ Red CMXRos for 20 minutes at 37°C. After incubation, cells were washed with Annexin V binding buffer twice and stained with 1.0 µg/T of Alexa Fluor® 488 Annexin V for ten minutes at room temperature.
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Description

MitoSpy™ mitochrondrial localization probes are cell-permeant, fluorogenic chemical reagents used for labeling mitochondria of living cells. MitoSpy™ Red CMXRos’ attraction to the mitochondria is not based on membrane potential and thus can be used to measure mitochondrial mass of individual cells in flow cytometry.

Product Details
Cat # Size Price Quantity Avail. Save
424801 5 x 50 µg $65.00
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424802 20 x 50 µg $210.00
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Technical data sheet

Product Details

Molecular Mass
531.52 g/mol
Preparation
The stock solution for MitoSpy™ Red CMXRos is prepared by dissolving the lyophilized probe in dimethyl sulfoxide (DMSO) to make a final concentration of 1mM by adding 94µl of DMSO to each vial.
Storage & Handling
Store MitoSpy™ Red CMXRos at -20°C.
Application

IF - Quality tested
FC - Validated

Recommended Usage

Each lot of this reagent is quality control tested by immunofluorescence staining. For immunofluorescence microscopy, a concentration range of 50 nM to 500 nM is recommended. For flow cytometric staining, a concentration range of 10 nM to 50 nM is recommended. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

MitoSpy™ Red CMXRos is excited at 577nm and emits at 598nm.

1. Reconstitute MitoSpy™ Red CMXRos to a 1mM concentration with DMSO by adding 94µl DMSO to an individual vial of lyophilized material. Protect the stock solution from light and keep frozen for storage.
2. Prepare the working solution for MitoSpy™ Red CMXRos in 37°C culture medium (incomplete), this will vary by cell line and type of imaging required.

  • If labeling mitochondria for live cell imaging a concentration between 50-250 nM is recommended.
  • If cells are labeled live and then subsequently fixed a concentration between 250-500nM recommended.

3. Grow cells to a desired confluency and wash once with warm 1X PBS.
4. Add the diluted MitoSpy™ Red CMXRos solution to the live cells and place them in a 37°C incubator for 20-30 minutes.
5. Wash the cells twice with warm 1X PBS or culture media.
6. If the cells will be imaged live, they can now be imaged with a fluorescence microscope.

If the cells need to be fixed:

1. Fix the cells with 2-4% paraformaldehyde (PFA) for 10 mins at room temperature.
2. Wash the cells twice with 1X PBS.
3. Regular IF staining protocol can be used for antibodies or other probe co-stains.

If the cells need to be permeabilized:

1. Dilute the 10X True Nuclear™ 10X Perm Buffer in DI water.
2. Incubate cells with 1X perm buffer for 10 minutes at room temperature.
3. Wash cells with 1X PBS twice.

Antigen Details

Distribution
Mitochondria.
Gene ID
NA

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Go To Top Version: 1    Revision Date: 12/19/2016

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