Biolegend, 424805, MitoSpy™ Green FM
5x50microg
Catalog Number: 424805
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| Verified Reactivity | Human, Mouse, Rat, All Species |
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| Molecular Mass | 671.88 g/mol |
| Preparation | The stock solution for MitoSpy™ Green FM is prepared by dissolving the lyophilized probe in dimethyl sulfoxide (DMSO) to make a final concentration of 1 mM by adding 74 µl of DMSO to each vial. |
| Storage & Handling | Store MitoSpy™ Green FM at -20°C. |
| Application | ICC - Quality tested |
| Recommended Usage | Each lot of this reagent is quality control tested by immunocytochemistry staining. For immunocytochemistry microscopy, a concentration range of 50nM to 500nM is recommended. It is recommended that the reagent be titrated for optimal performance for each application. |
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| Application Notes |
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| MitoSpy™ Green FM is excited at 490nm and emits at 516 nm. 1. Prior to reconstitution, spin down the vial of lyophilized reagent in a microcentrofuge to ensure the reagent is at the bottom of the vial. 2. Reconstitute MitoSpy™ Green FM to a 1 mM concentration with DMSO by adding 74 µl DMSO to an individual vial of lyophilized material. Protect the stock solution from light and keep frozen for storage. 3. Prepare the working solution for MitoSpy™ Green FM 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 - 500 nM is recommended. 4. Grow cells to a desired confluency and wash once with warm 1 x PBS. 5. Add the diluted MitoSpy™ Green FM solution to the live cells and place in the 37°C incubator for 20-30 minutes. 6. Wash the cells twice with warm 1 X PBS or culture media. 7. If the cells will be imaged live, they can now be imaged with a fluorescence microscope. If the cells need to be fixed: A. Fix the cells with 2-4% paraformaldehyde (PFA) for ten minutes at room temperature. B. Wash the cells twice with 1 x PBS. C. Regular IF staining protocol can be used for antibodies or other probe co-stains. |
| Product Citations | Galli A, et al. 2020. Biomedicines. :8. PubMed Maffioli E, et al. 2020. Front Cell Dev Biol. 508:8. PubMed Dent JR, et al. 2019. J Appl Physiol (1985). 127:1117. PubMed |
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| Distribution | Mitochondria. |
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| Biology Area | Apoptosis/Tumor Suppressors/Cell Death, Cell Biology, Mitochondrial Function, Neuroscience |
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| Molecular Family | Mitochondrial Markers |
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| Gene ID | NA |
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| Regulatory Status | RUO |
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| Other Names | Mitochondrial labeling |
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